LOCUS       40924_1044        6910 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Conditional gene trap vector GBT-S8, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  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 6910)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6910)
  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..6910
                     /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(2150..2183)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     protein_bind    complement(2190..2223)
                     /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)."
     polyA_signal    2358..2492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2504..3220)
                     /codon_start=1
                     /label=BFP
                     /note="blue variant of GFP"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     promoter        complement(3983..4001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4008..4024)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4165..4620
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4698..4802
                     /label=AmpR promoter
     CDS             4803..5660
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5834..6422
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6710..6731
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6746..6776
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6800
                     /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     6808..6824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6842..6860
                     /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 tcgacgcggc cgctctagag aagttcatat
     2161 actttctaga gaataggaac ttcgctagct accgttcgta tagcatacat tatacgaagt
     2221 tatattaccc tgttatccct agatctattt aaatgataat cagccatacc acatttgtag
     2281 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2341 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2461 aactcatcaa tgtatcttat catgtctgga tcatcatcga ttacttgtac agctcgtcca
     2521 tgccgtgagt gatgccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2581 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
     2641 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     2701 cctcaatatt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcca
     2761 tgatatagac gttgtggctg ttgaagttgt actcgagctt gtgccccagg atgttgccgt
     2821 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2881 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2941 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3001 ggctgaagca ctgcacgccg tgggacaggg tggtcactag tgtgggccag ggcacgggca
     3061 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3121 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     3181 gcaccacccc ggtgaacagc tcctcgccct tgctagccat ggtggatccc ccgcaagtct
     3241 gactgagtgg aagtttcagt tcaatggaag ggagaaaaga gccagccttt atatatgtca
     3301 tgtattgctg ctgtctgcat gcgatgaacc tagagcttac tttttctgtc aagtaaggca
     3361 aaagcagaga acctttgtct tgcattgtgt ttgatgttgt gctgtgacag tcatttctta
     3421 gttcatgctg gtacttttgt tccatagctt tacccaaaga gttatccagc attcccagtt
     3481 attccaaaaa ctcaatgtca caaattttcc agtccattgc ttagcagcgt tgaccccgga
     3541 gaccctgatg tggtaattta cagtgcaatt cagttgctgg tccaaacact ttgtaacagt
     3601 ttaaagaatt gtggttacta gtaaatcctt aatttggcat catctaggtg tcattctttg
     3661 ccacttttga atcacacaaa tgaaccaaac aatggttagc tagcagatca aattaaactg
     3721 ggcatcagcg caattcaatt ggtttggtaa tagcaaggga aaatagaatg aagtgatctc
     3781 caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag tacttttttt
     3841 tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta ctcaagtaaa
     3901 gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac tcaagtactt
     3961 tacacctctg ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     4021 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4081 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4141 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4201 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4261 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4321 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4381 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4441 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4501 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4561 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4621 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tcaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6541 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6601 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6661 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6721 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6781 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6841 tatttaggtg acactataga atactcaagc tatgcatcca acgcgttggg agctctccca
     6901 tatggtcgag
//
