LOCUS       40924_1039        6655 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Conditional gene trap vector GBT-S1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   Grajevskaja V, Camerota D, Bellipanni G, Balciuniene J, Balciunas D.
  TITLE     Analysis of a conditional gene trap reveals that tbx5a is required 
            for heart regeneration in zebrafish
  JOURNAL   PLoS ONE 13 (6), e0197293 (2018)
  PUBMED    29933372
REFERENCE   2  (bases 1 to 6655)
  AUTHORS   Camerota D, Balciunas D.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2018) Department of Biology, Temple University, 
            1900 N 12th St, Philadelphia, PA 19122, USA
REFERENCE   3  (bases 1 to 6655)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2018"; volume: "13"; issue: "6"; pages: "e0197293"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2018) Department of Biology, Temple University, 1900 N 12th 
            St, Philadelphia, PA 19122, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    220..253
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    259..292
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             469..906
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    complement(2151..2184)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     protein_bind    complement(2191..2224)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2485..3201
                     /codon_start=1
                     /label=BFP
                     /note="blue variant of GFP"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     polyA_signal    complement(3213..3347)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3728..3746)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3753..3769)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3910..4365
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4443..4547
                     /label=AmpR promoter
     CDS             4548..5405
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5579..6167
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6455..6476
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6491..6521
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6529..6545
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6553..6569
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6587..6605
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa
       61 attttactca attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa
      121 agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa
      181 actagagatt cttgtttaag cttagggata acagggtaat accgttcgta taatgtatgc
      241 tatacgaagt tatctgcaga agttcatatt ctctagaaag tataggaact tccatggaat
      301 tcactagtca cccactagcg gtcagactgc agattgcagc acgaaacagg aagctgactc
      361 cacatggtca catgctcact gaagtgttga cttccctgac agctgtgcac tttctaaacc
      421 ggttttctca ttcatttaca gttcagccgg aattcacccg ggtaccagaa gctactgtct
      481 tctatcgaac aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa
      541 ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa
      601 aggtctccgc tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa
      661 cagctatttc tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct
      721 ttacaggata taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat
      781 gccgtcacag atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat
      841 agaataagtg cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact
      901 gtatcgtcga ggtcgacccc gggaattcag atctctcgag ccgccccccc gaccgatgtc
      961 agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg
     1021 ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc
     1081 ccccacgact ccgcccccta cggcgctctg gatatgtaaa tcgatacgga ctgttaccac
     1141 ttcacgccga ctcaaactgc gcagagaaac ttcaaacgac aacattggca tggcttttgt
     1201 tatttttggc gcttgactca ggatctaaaa actggaacgg tgaaggtgac ggcaatgttt
     1261 tttggcaaat aagcatcccc gaagttctac aatgcatctg aggactccat gttttgtttt
     1321 ctttagtcat tccaaatgtt tgttaaatgc attgttccga aacttatttg cctctatgaa
     1381 ggctgcccag taattgggag catacttaac attgtagtat tgtatgtaaa ttatgtaaca
     1441 aaacaatgtc tgggtttttg tacttttcag ccttaaaatc ttgggtttat ttttttttct
     1501 ttgttccaaa aaaacgaagc tttaccattc aagatgtaaa ggtttccatt ccccctgggc
     1561 atattgtaaa agctgtgtgg aacgtggcgg tgccagacat ttggtggggc caacctgtac
     1621 actgactaat tcaattccaa taaaagtgca catgtaagac atcctacccc tgtctcattt
     1681 cctgtcttgg ctctgagtgg cttcaataaa aatggctttt gtgaagttgc accatgttaa
     1741 ttacatttca gcactttcac accaggtttt tgactaattt gaatttatcg aagtttgacc
     1801 tttttttttt tttttccttc aagctaattt gaggttatca aaccatttca ttggaggaaa
     1861 cttaatgttt aaaagcataa attcgaaaca tgattcattt atttaactgc ttaatggtga
     1921 aacaaacttc aagtaataca ccaagtgttc cctcatatga aggtagctaa ctgcaaacgg
     1981 ccttaacttt ccatcttaag ttgagctaca taaaggcttt aacaggaagg aagagccagt
     2041 ttaaaagtca aaccaccatg actgatataa attcttgtcc atataacctt gcgtttcaca
     2101 caatatagct atctactgct aaataaatta tcgcgagcgg ccgctctaga gaagttcata
     2161 tactttctag agaataggaa cttcgctagc taccgttcgt atagcataca ttatacgaag
     2221 ttatattacc ctgttatccc tagatctatt tgatctgcta gcgaacaaaa gtaccagcat
     2281 gaactaagaa atgactgtca cagcacaaca tcaaacacaa tgcaagacaa aggttctctg
     2341 cttttgcctt acttgacaga aaaagtaagc tctaggttca tcgcatgcag acagcagcaa
     2401 tacatgacat atataaaggc tggctctttt ctcccttcca ttgaactgaa acttccactc
     2461 agtcagactt gcgggggatc caccatggct agcaagggcg aggagctgtt caccggggtg
     2521 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2581 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2641 aagctgcccg tgccctggcc cacactagtg accaccctgt cccacggcgt gcagtgcttc
     2701 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2761 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2821 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2881 gaggacggca acatcctggg gcacaagctc gagtacaact tcaacagcca caacgtctat
     2941 atcatggccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaatatt
     3001 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3061 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3121 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg catcactcac
     3181 ggcatggacg agctgtacaa gtaatcgatg atgatccaga catgataaga tacattgatg
     3241 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3301 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3361 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3421 acctctacaa atgtggtatg gctgattatc atttaaatta aactgggcat cagcgcaatt
     3481 caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact
     3541 ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa
     3601 ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa
     3661 aataatactt aagtacagta atcaagtaaa attactcaag tactttacac ctctgggccc
     3721 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     3781 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3841 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3901 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4321 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     4381 tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg
     4441 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4501 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4561 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4621 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4681 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4741 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4801 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4861 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4921 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4981 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5041 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5101 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5161 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5221 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5281 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5341 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5401 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5461 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5521 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5701 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6241 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6301 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6361 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6421 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6481 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6541 aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact
     6601 atagaatact caagctatgc atccaacgcg ttgggagctc tcccatatgg tcgag
//
