LOCUS       Exported                6506 bp ds-DNA     circular SYN 05-DEC-2022
DEFINITION  Barcoded piggyBac self-reporting transposon with tdTomato marker in 
            AAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-PB-SRT-tdTomato_BC18
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6506
                     /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   4406..4535
                     /label=AAV2 ITR
     primer_bind     complement(4663..4680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4717..4760
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4817..5405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4897..4916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5576..6436)
                     /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     6199..6218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6437..6500)
                     /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 ggaaggttaa gagctcatct gagatgcagg tacatcccac tgatgagtcc
     4141 caaataggac gaaacgcgct tcggtgcgtc tgggattcca ctgctatcca cgcggccgca
     4201 cggcattacg ttagacgtct aggccacacc accctgaagg cgcctgctcg cctctgatct
     4261 gttgaagcta agcagggtcg gtcctggtta gtacttggat gggactccgc ctggtaatag
     4321 ccggtaccgt aggcttgcat ggtcatagct gtttccttgc gctggggatg cggtgggctc
     4381 tatagagagt cgatcgtgtg atcgaaggaa cccctagtga tggagttggc cactccctct
     4441 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4501 gcccgggcgg cctcagtgag cgagcgagcg cgcagcatgt agagactggg cctcatgggc
     4561 cttccgccta tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca
     4621 tggtcatagc tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct
     4681 gcgctcggtc gttcgggtaa agcctggggt gcctaaataa aacgaaaggc ccagtcttcc
     4741 gactgagcct ttcgttttat tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5221 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5521 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5641 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5701 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     5761 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6001 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6241 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6421 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6481 gtctcatgag cggatacata tttgaa
//
