LOCUS       Cellecta-pRSITPG        8890 bp ds-DNA     circular SYN 17-MAY-2019
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8890)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 17, 2019 from SnapGene 4.3.7 to Vector NTI(R) 
            format
            https://www.snapgene.com
COMMENT     VNTDATE|309830400|
COMMENT     VNTDBDATE|877910400|
COMMENT     VNTNAME|Cellecta-pRSITPGP-U6Tet-sh-HTS3-PGK-TetRep-2A-TagGFP2-2A-Pur
            o.dna|
COMMENT     VNTAUTHORNAME|Cellecta|
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..2064
                     /label=U6T8T
                     /label=U6-Tet
     protein_bind    2019..2037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2046..2064
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2066..2091
                     /label=cloning site
     misc_feature    2195..2312
                     /label=cPPT
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2365..2865
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2878..3498
                     /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             3505..3558
                     /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             3565..4278
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
     CDS             4297..4350
                     /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             4357..4956
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    4969..5557
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5631..5864
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5936..6057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6097..6232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(6667..7255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7426..8286)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(8287..8391)
                     /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 atggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga
     1861 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     1921 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     1981 tttaaaatgg actatcatat gcttaccgta acttgaaagc cctatcagtg atagagattt
     2041 atatatccct atcagtgata gagacaccgg agtcttcttt tttgaagaca cttcgtgcgt
     2101 ttggggtttc ggactgtaga actctgaacc tctcggtggt cgccgtatca ttagaattct
     2161 cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2221 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2281 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2341 cactttggcc gccgttcgaa ggggttgcgc cttttccaag gcagccctgg gtttgcgcag
     2401 ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc
     2461 acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc
     2521 ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc
     2581 cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag
     2641 ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg
     2701 cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg
     2761 tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg
     2821 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagggtct agacaccatg
     2881 tccagactgg acaagagcaa agtcatcaac tctgccttgg agctcctgaa tgaagttggc
     2941 attgagggct tgaccaccag gaagctggcc cagaagctgg gtgtggagca gcctaccctg
     3001 tactggcatg tgaagaacaa gagggctctg cttgatgccc tggccattga gatgttggac
     3061 aggcaccaca cccacttctg ccctctggaa ggggagtcct ggcaggactt cctgaggaac
     3121 aatgccaaga gcttcagatg tgccttgctc tcccaccggg acggtgccaa agttcacttg
     3181 ggcaccaggc ctacagagaa gcagtatgag accctggaga accagctggc attcctgtgc
     3241 caacaaggct tctccctgga aaatgccttg tatgccctct ctgctgtggg ccacttcacc
     3301 ttgggctgtg tgctggagga ccaggagcac caagttgcca aggaggagag ggagaccccc
     3361 accactgact ccatgccacc actgctgcgg caagctattg agttgtttga ccaccaaggg
     3421 gctgagcctg cattcctgtt tggcctggaa ctgatcatct gtggcctgga aaagcagctg
     3481 aaatgtgagt ctggctctgg atccgagggc agaggaagcc ttctaacatg cggtgacgtg
     3541 gaggagaatc ccggcccttc cggaatgagc gggggcgagg agctgttcgc cggcatcgtg
     3601 cccgtgctga tcgagctgga cggcgacgtg cacggccaca agttcagcgt gcgcggcgag
     3661 ggcgagggcg acgccgacta cggcaagctg gagatcaagt tcatctgcac caccggcaag
     3721 ctgcccgtgc cctggcccac cctggtgacc accctctgct acggcatcca gtgcttcgcc
     3781 cgctaccccg agcacatgaa gatgaacgac ttcttcaaga gcgccatgcc cgagggctac
     3841 atccaggagc gcaccatcca gttccaggac gacggcaagt acaagacccg cggcgaggtg
     3901 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcaagga cttcaaggag
     3961 gacggcaaca tcctgggcca caagctggag tacagcttca acagccacaa cgtgtacatc
     4021 cgccccgaca aggccaacaa cggcctggag gctaacttca agacccgcca caacatcgag
     4081 ggcggcggcg tgcagctggc cgaccactac cagaccaacg tgcccctggg cgacggcccc
     4141 gtgctgatcc ccatcaacca ctacctgagc actcagacca agatcagcaa ggaccgcaac
     4201 gaggcccgcg accacatggt gctcctggag tccttcagcg cctgctgcca cacccacggc
     4261 atggacgagc tgtacagggc tagcaccggt agatccgagg gcagaggaag ccttctaaca
     4321 tgcggtgacg tggaggagaa tcccggccct tccgggatga ccgagtacaa gcccacggtg
     4381 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4441 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     4501 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     4561 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     4621 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     4681 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     4741 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     4801 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     4861 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4921 cgcacctggt gcatgacccg caagcccggt gcctgaatct aggtcgacaa tcaacctctg
     4981 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5041 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5101 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5161 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5221 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5281 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5341 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5401 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5461 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtctccg ccttcgccct
     5521 cagacgagtc ggatctctct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     5581 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5641 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga
     5701 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5761 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5821 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5881 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5941 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6001 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6061 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6181 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6241 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     6301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7081 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8401 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8461 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8521 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8581 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8641 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     8701 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     8761 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8821 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     8881 tgccaagctg
//
