LOCUS       Exported                6504 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  piggyBac self-reporting transposon with tdTomato marker in AAV 
            backbone.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-PB-SRT-tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  AUTHORS   Cammack AJ, Moudgil A, Chen J, Vasek MJ, Shabsovich M, McCullough K,
            Yen A, Lagunas T, Maloney SE, He J, Chen X, Hooda M, Wilkinson MN, 
            Miller TM, Mitra RD, Dougherty JD
  TITLE     A viral toolkit for recording transcription factor-DNA interactions 
            in live mouse tissues.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 5;117(18):10003-10014. doi: 
            10.1073/pnas.1918241117. Epub 2020 Apr 16.
  PUBMED    32300008
REFERENCE   2  (bases 1 to 6504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   44..173
                     /label=AAV2 ITR
     repeat_region   320..382
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        625..1806
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          855..1797
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1754..1774
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1818..3248
                     /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   3630..3664
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   3972..4101
                     /label=AAV2 ITR
     primer_bind     complement(4229..4246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4283..4326
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4383..4971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4463..4482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5142..6002)
                     /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     5765..5784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6003..6066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6451..6473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6465..6482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6466..6482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6493..8
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6493..7
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctatagggcg aattgtaggc ggaaggccgt caaggccgca tggctgcgcg ctcgctcgct
       61 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctattgacg
      181 tcagatcgct aaataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 acgtcaaggc cgcatttaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa
      361 tcatgtgtaa aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat
      421 ttgaatagat attaagtttt attatattta cacttacata ctaataataa attcaacaaa
      481 caatttattt atgtttattt atttattaaa aaaaacaaaa actcaaaatt tcttctataa
      541 agtaacaaaa cttttatcga attcctgcag cccagcttga tcttaggtcc ctcgacgaat
      601 tctctagaga agagcatgcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
      661 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
      721 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      781 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      841 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
      901 atggcccttg cgtgccttga attacttcca cgcccctggc tgcagtacgt gattcttgat
      961 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
     1021 tcgcctcgtg cttgagttga ggcctggctt gggcgctggg gccgccgcgt gcgaatctgg
     1081 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
     1141 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
     1201 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
     1261 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
     1321 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     1381 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     1441 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     1501 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     1561 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     1621 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     1681 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     1741 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     1801 tcgtgaaaac tacgccaatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct
     1861 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1921 agggccgccc ctacgagggc acccaggccg ccaagctgaa ggtgaccaag ggcggccccc
     1981 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     2041 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     2101 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     2161 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     2221 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2281 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     2341 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2401 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2461 acgagcgctc cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca
     2521 gcggcagctc cggcaccgcc tcctccgagg acaacaacat ggccgtcatc aaagagttca
     2581 tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     2641 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     2701 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt
     2761 acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca
     2821 agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct
     2881 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     2941 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     3001 cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc
     3061 actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     3121 actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     3181 aacagtacga gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt
     3241 acaagtaatc gataacaact cgagtaccgg cttaagtgat agatcggaag agcggttcag
     3301 caggaatgcc gagaccggtt taaactcagt gtccatagcg acgccggcgc gataagctat
     3361 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
     3421 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg
     3481 actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc
     3541 acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
     3601 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
     3661 agggttaaga gctcctgaga tgcaggtaca tcccactgat gagtcccaaa taggacgaaa
     3721 cgcgcttcgg tgcgtctggg attccactgc tatccacgcg gccgcacggc attacgttag
     3781 acgtctaggc cacaccaccc tgaaggcgcc tgctcgcctc tgatctgttg aagctaagca
     3841 gggtcggtcc tggttagtac ttggatggga ctccgcctgg taatagccgg taccgtaggc
     3901 ttgcatggtc atagctgttt ccttgcgctg gggatgcggt gggctctata gagagtcgat
     3961 cgtgtgatcg aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4021 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4081 agtgagcgag cgagcgcgca gcatgtagag actgggcctc atgggccttc cgcctatcac
     4141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt
     4201 tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4261 gggtaaagcc tggggtgcct aaataaaacg aaaggcccag tcttccgact gagcctttcg
     4321 ttttattgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4801 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5161 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5221 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5281 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5341 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5401 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5461 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5521 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5581 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5641 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5701 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5761 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5821 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5881 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5941 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6001 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6061 tacatatttg aagctaataa caggcctgct ggtaatcgca ggccttttta tttggtgtat
     6121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     6181 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6241 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6301 ttgagtggcc gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg
     6361 gcgtttcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6421 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6481 gtgagcgcga cgtaatacga ctca
//
