LOCUS       Exported                6510 bp ds-DNA     circular SYN 06-DEC-2022
DEFINITION  Barcoded piggyBac self-reporting transposon with tdTomato marker in 
            AAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-PB-SRT-tdTomato_BC6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   Lalli M, Yen A, Thopte U, Dong F, Moudgil A, Chen X, Milbrandt J, 
            Dougherty JD, Mitra RD
  TITLE     Measuring transcription factor binding and gene expression using 
            barcoded self-reporting transposon calling cards and transcriptomes.
  JOURNAL   NAR Genom Bioinform. 2022 Aug 31;4(3):lqac061. doi: 
            10.1093/nargab/lqac061. eCollection 2022 Sep.
  PUBMED    36062164
REFERENCE   2  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 6, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6510
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     379..401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     393..410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     394..410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     421..440
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        421..439
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   476..605
                     /label=AAV2 ITR
     repeat_region   752..814
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1057..2238
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1287..2229
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2186..2206
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2250..3680
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQAA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     repeat_region   4410..4539
                     /label=AAV2 ITR
     primer_bind     complement(4667..4684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4721..4764
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4821..5409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4901..4920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5580..6440)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6203..6222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6441..6504)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctaataaca ggcctgctgg taatcgcagg cctttttatt tggtgtattt agaaaaataa
       61 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
      121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
      241 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
      301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
      421 taatacgact cactataggg cgaattgtag gcggaaggcc gtcaaggccg catggctgcg
      481 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      541 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      601 ttcctattga cgtcagatcg ctaaataatg acgtatgttc ccatagtaac gccaataggg
      661 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      721 caagtgtatc atacgtcaag gccgcattta accctagaaa gataatcata ttgtgacgta
      781 cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
      841 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
      901 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa
      961 tttcttctat aaagtaacaa aacttttatc gaattcctgc agcccagctt gatcttaggt
     1021 ccctcgacga attctctaga gaagagcatg cgtgaggctc cggtgcccgt cagtgggcag
     1081 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     1141 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     1201 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1261 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     1321 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacgcccctg gctgcagtac
     1381 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     1441 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ttgggcgctg gggccgccgc
     1501 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     1561 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     1621 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     1681 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     1741 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     1801 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     1861 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     1921 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1981 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     2041 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     2101 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     2161 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     2221 ccatttcagg tgtcgtgaaa actacgccaa tggtgagcaa gggcgaggag gtcatcaaag
     2281 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     2341 agggcgaggg cgagggccgc ccctacgagg gcacccaggc cgccaagctg aaggtgacca
     2401 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     2461 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     2521 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     2581 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     2641 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     2701 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     2761 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     2821 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     2881 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg catggcaccg
     2941 gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac atggccgtca
     3001 tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg
     3061 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     3121 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg
     3181 gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc
     3241 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga
     3301 cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca
     3361 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg
     3421 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga
     3481 aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc
     3541 aactgcccgg ctactactac gtggacacca agctggacat cacctcccac aacgaggact
     3601 acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtacggca
     3661 tggacgagct gtacaagtaa tcgataacaa ctcgagtacc ggcttaagtg atagatcgga
     3721 agagcggttc agcaggaatg ccgagaccgg tttaaactca gtgtccatag cgacgccggc
     3781 gcgataagct atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     3841 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     3901 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     3961 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     4021 attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca
     4081 gactatcttt gtcaggttaa gagctcaagc ttctgagatg caggtacatc ccactgatga
     4141 gtcccaaata ggacgaaacg cgcttcggtg cgtctgggat tccactgcta tccacgcggc
     4201 cgcacggcat tacgttagac gtctaggcca caccaccctg aaggcgcctg ctcgcctctg
     4261 atctgttgaa gctaagcagg gtcggtcctg gttagtactt ggatgggact ccgcctggta
     4321 atagccggta ccgtaggctt gcatggtcat agctgtttcc ttgcgctggg gatgcggtgg
     4381 gctctataga gagtcgatcg tgtgatcgaa ggaaccccta gtgatggagt tggccactcc
     4441 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4501 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc atgtagagac tgggcctcat
     4561 gggccttccg cctatcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact
     4681 cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa ataaaacgaa aggcccagtc
     4741 ttccgactga gcctttcgtt ttattgagca aaaggccagc aaaaggccag gaaccgtaaa
     4801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5641 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5701 agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac
     5761 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5821 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5881 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5941 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6001 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6061 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6121 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6181 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6241 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6301 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6361 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6421 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6481 tattgtctca tgagcggata catatttgaa
//
