LOCUS       Cellecta-pRSITH1        8843 bp ds-DNA     circular SYN 13-SEP-2019
DEFINITION  pRSITH1PRP-H1Tet-sh-HTS3-PGK-TetRep-2A-TagRFP-2A-Puro.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8843)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 13, 2019 from SnapGene 4.3.11 to Vector NTI(R) 
            format
            https://www.snapgene.com
COMMENT     VNTDATE|712368000|
COMMENT     VNTDBDATE|888624000|
COMMENT     VNTNAME|Cellecta-pRSITH1PRP-H1Tet-sh-HTS3-PGK-TetRep-2A-TagRFP-2A-Pu
            ro|
FEATURES             Location/Qualifiers
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        1803..2021
                     /label=H1-Tet
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    1975..1993
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2003..2021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2022..2047
                     /label=cloning site
     misc_feature    2151..2268
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2321..2821
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2834..3454
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
     CDS             3461..3514
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
     CDS             3521..4231
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
     CDS             4250..4303
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
     CDS             4310..4909
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    4922..5510
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5584..5817
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5889..6010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6050..6185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(6620..7208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7379..8239)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(8240..8344)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 atgaacgctg acgtcatcaa cccgctccaa ggaatcgcgg gcccagtgtc actaggcggg
     1861 aacacccagc gcgcgtgcgc cctggcagga agatggctgt gagggacagg ggagtggcgc
     1921 cctgcaatat ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt gaaatcccta
     1981 tcagtgatag agacttataa gttccctatc agtgatagag aaccggagtc ttcttttttg
     2041 aagacacttc gtgcgtttgg ggtttcggac tgtagaactc tgaacctctc ggtggtcgcc
     2101 gtatcattag aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     2161 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2221 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     2281 agggacagca gagatccact ttggccgccg ttcgaagggg ttgcgccttt tccaaggcag
     2341 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     2401 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     2461 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     2521 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     2581 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     2641 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     2701 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     2761 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     2821 gggtctagac accatgtcca gactggacaa gagcaaagtc atcaactctg ccttggagct
     2881 cctgaatgaa gttggcattg agggcttgac caccaggaag ctggcccaga agctgggtgt
     2941 ggagcagcct accctgtact ggcatgtgaa gaacaagagg gctctgcttg atgccctggc
     3001 cattgagatg ttggacaggc accacaccca cttctgccct ctggaagggg agtcctggca
     3061 ggacttcctg aggaacaatg ccaagagctt cagatgtgcc ttgctctccc accgggacgg
     3121 tgccaaagtt cacttgggca ccaggcctac agagaagcag tatgagaccc tggagaacca
     3181 gctggcattc ctgtgccaac aaggcttctc cctggaaaat gccttgtatg ccctctctgc
     3241 tgtgggccac ttcaccttgg gctgtgtgct ggaggaccag gagcaccaag ttgccaagga
     3301 ggagagggag acccccacca ctgactccat gccaccactg ctgcggcaag ctattgagtt
     3361 gtttgaccac caaggggctg agcctgcatt cctgtttggc ctggaactga tcatctgtgg
     3421 cctggaaaag cagctgaaat gtgagtctgg ctctttcgaa gagggcagag gaagccttct
     3481 aacatgcggt gacgtggagg agaatcccgg cccttccgga atggtgtcta agggcgaaga
     3541 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca
     3601 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     3661 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     3721 gtacggcagc agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     3781 cttccctgag ggcttcacat gggagagagt caccacatac gaggacgggg gcgtgctgac
     3841 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     3901 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccaa
     3961 caccgagatg ctgtaccccg ctgacggcgg cctggaaggc agaagcgaca tggccctgaa
     4021 gctcgtgggc gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc
     4081 cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac cacagactgg aaagaatcaa
     4141 ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga
     4201 cctccctagc aaactggggc acaaacttaa tgctagcacc ggtagatccg agggcagagg
     4261 aagccttcta acatgcggtg acgtggagga gaatcccggc ccttccggga tgaccgagta
     4321 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4381 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4441 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4501 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4561 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4621 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4681 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4741 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4801 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4861 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa tctaggtcga
     4921 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4981 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5041 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5101 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5161 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5221 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5281 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5341 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5401 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5461 ccgccttcgc cctcagacga gtcggatctc tctttgggcc gcctccccgc ctggtacctt
     5521 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5581 gactggaagg gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt
     5641 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5701 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5761 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     5821 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     5881 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5941 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6001 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     6061 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6121 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6181 aggcctagac ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa
     6241 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6301 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6361 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6421 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6481 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6541 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6601 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6661 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6721 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6781 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6841 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6901 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6961 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7021 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7081 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7141 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7201 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7261 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7321 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7381 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7441 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7501 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7561 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7621 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7681 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7741 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7801 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7861 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7921 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7981 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8041 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8101 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8161 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8221 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8281 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8341 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8401 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     8461 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     8521 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     8581 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     8641 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     8701 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     8761 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     8821 aaacgacggc cagtgccaag ctg
//
