LOCUS       Cellecta-pScribe        7495 bp ds-DNA     circular SYN 12-FEB-2020
DEFINITION  pScribe4-EFS-TagRFP-2A-Puro-bc-3LTR barcode expression vector 
            showing amplification and sequencing primers for NGS of DNA for the 
            CloneTracker XP Barcode Libraries. Cloned barcode 
            BC14.001-BC30.010000 is shown.The primer for scRNA-Seq of barcodes 
            on the 10x Genomics platform is also shown (Adaptor-FBP1). If you 
            are running other single cell bead-based platforms (e.g., Illumina 
            Bio-Rad SureCell), you will need to modify the adaptor portion of 
            the primer based on the sequences used in the single-cell protocols 
            for those systems.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7495)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Feb 12, 2020 from SnapGene 5.0.7 to Vector 
            NTI(R) format
            https://www.snapgene.com
COMMENT     For library sample multiplexing (up to 24 samples per lane), please 
            use Cellecta's NGS Prep Kit for Barcode Libraries in pScribe 
            (CloneTracker XP), Cat.# LNGS-300, and Supplementary Primer Set, 
            Cat.# LNGS-300-SP.
COMMENT     VNTDATE|840672000|
COMMENT     VNTDBDATE|905126400|
COMMENT     VNTNAME|Cellecta-pScribe4-EFS-TagRFP-2A-Puro-bc-3LTR|
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."
     misc_feature    1807..1922
                     /label=cPPT
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        1931..2141
                     /label=EFS
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2154..2864
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
     CDS             2871..2924
                     /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             2931..3530
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    3540..4128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_feature    4137..4235
                     /label=3LTR (truncated)
     primer          4252..4276
                     /label=FSeqRNA-BC14-XP Primer (Adaptor-FBP1)
                     /note="sequence: 
                     GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTCCGACCACCGAACGCAACGCACGCA
                     "
     primer          4257..4276
                     /label=NFwd-XP Primer
                     /note="sequence: 
                     AGCAGAAGACGGCATACGAGATAGCACCGAACGCAACGCACGCA"
     primer          4258..4276
                     /label=Forward-XP Primer
                     /note="sequence: ACCGAACGCAACGCACGCA"
     misc_feature    4277..4324
                     /label=BC14 + 4 + BC30
     primer          complement(4325..4348)
                     /label=Seq-XP NGS Primer
                     /note="sequence: AGACGACCTGCTCCAGCTGCACCA"
     primer          complement(4383..4414)
                     /label=NRev-XP Primer
                     /note="sequence: 
                     AGATACGGCGACCACCGAGATCTACACGCACGACGAGACGCAGACGAATACGACACGAC
                     GACCGACCCGAACCACGA"
     primer          complement(4383..4408)
                     /label=Reverse-XP Primer
                     /note="sequence: ACGACCACGACCGACCCGAACCACGA"
     LTR             4420..4634
                     /label=3' LTR
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    4691..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4852..4987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5422..6010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6181..7041)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(7042..7146)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7194..7328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
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 attcaattta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     1861 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     1921 ttctcgaggt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     1981 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2041 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2101 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac atctagagcc accatggtgt
     2161 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     2221 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     2281 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     2341 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
     2401 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaggacg
     2461 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2521 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2581 gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     2641 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
     2701 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
     2761 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     2821 ccagatactg cgacctccct agcaaactgg ggcacaaact taatggatcc gagggcagag
     2881 gaagccttct aacatgcggt gacgtggagg agaatcccgg cccttccgga atgaccgagt
     2941 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3001 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     3061 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     3121 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     3181 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     3241 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     3301 tcctggccac cgtcggcgtg tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     3361 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     3421 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     3481 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctaa tgagtcgaca
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcctc
     4081 gccttcgccc tcagacgagt cggatctctc tttgggccgc ctccccgcct ggtaccttta
     4141 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     4201 ctggaagggc taattcactc ccaacgaaga taagattcga agaattcacg accgaccacc
     4261 gaacgcaacg cacgcagttt agatatacac tggtgtgtgt gtgttggttg caacacacac
     4321 gtcatggtgc agctggagca ggtcgtctgt cgtggtagtg atgtggtatc tggtgagtac
     4381 tgtcgtggtt cgggtcggtc gtggtcgtga attcaccggt ctgctttttg cttgtactgg
     4441 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     4501 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     4561 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4621 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     4681 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4801 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     4861 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4921 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4981 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     5041 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5101 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5161 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5221 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5281 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5341 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5401 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5461 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5521 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5581 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5641 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5701 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5761 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5821 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5881 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5941 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6001 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6061 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6121 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6181 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6241 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6301 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6361 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6421 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6481 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6541 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6601 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6661 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6721 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6781 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6841 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6901 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6961 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7021 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7081 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7141 tccgcgcaca tttccccgaa aagtgccacc tgacgtcaac ggatcgggag atcaacttgt
     7201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7321 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
     7381 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
     7441 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
