LOCUS       sequence-182581-        4329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry TOPO TA cloning vector for SRF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4329)
  AUTHORS   Bouveret R, Waardenberg AJ, Schonrock N, Ramialison M, Doan T, de 
            Jong D, Bondue A, Kaur G, Mohamed S, Fonoudi H, Chen CM, Wouters MA,
            Bhattacharya S, Plachta N, Dunwoodie SL, Chapman G, Blanpain C, 
            Harvey RP
  TITLE     NKX2-5 mutations causative for congenital heart disease retain 
            functionality and are directed to hundreds of targets.
  JOURNAL   Elife. 2015 Jul 6;4. doi: 10.7554/eLife.06942.
  PUBMED    26146939
REFERENCE   2  (bases 1 to 4329)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-07-6"; volume: "4. doi"; pages: " 10.7554/eLife.06942"
COMMENT     Sequence Label: pCR8/GW/TOPO-SRF
FEATURES             Location/Qualifiers
     source          1..4329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1672..1771)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1788..1807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1789..1807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1812..1828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1812..1828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2259..3050
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3143..3731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3632..3651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3885..3902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4061..4088)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4180..4266)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4315..4329,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4329,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgaattc gcccttacca tgttaccgag ccaagctggg gccgcggcgg
      181 ctctgggccg gggctcggcc ctggggggca acctgaaccg gaccccgacg gggcggccgg
      241 gcggcggcgg cgggactcgc ggggcgaacg ggggccgggt ccccgggaac ggcgcggggc
      301 tcggccagag tcgtctggag cgggaggctg cagcggcagc ggcgcccacc gccggggccc
      361 tctacagcgg cagcgagggc gactccgagt ccggcgagga ggaggagctg ggcgccgagc
      421 ggcgcggcct caagcggagc ctgagcgaga tggagctcgg cgtggtggtc ggtgggcctg
      481 aggcggcggc ggccgccggg ggctacgggc cggtgagcgg cgcggtgagc ggggccaagc
      541 cggggaagaa gacccggggc cgcgtgaaga tcaagatgga gttcatcgac aacaagctgc
      601 ggcgctacac gaccttcagc aagaggaaga cgggcatcat gaagaaggcc tatgagctgt
      661 ccacgctgac agggacacag gtgctgttgc tggtggccag tgagacaggc catgtgtata
      721 cctttgccac ccgcaaactg cagcccatga tcaccagtga gaccggcaag gcgctgattc
      781 agacctgcct caactcgcca gactctccgc cccgctcaga ccccaccaca gaccagagaa
      841 tgagtgccac tggctttgaa gagccagatc tcacctacca ggtgtcggaa tctgacagca
      901 gtggggaaac caaggacaca ctgaagccag cattcacagt caccaacctg ccgggtacca
      961 cctccacaat ccagacagca cccagcacct ctaccaccat gcaagtcagc agcggcccct
     1021 ccttccccat caccaactac ctggcaccag tgtctgctag tgtcagcccc agcgctgtca
     1081 gcagtgccaa cgggactgtg ctcaagagta caggcagcgg ccctgtctcc tctgggggcc
     1141 ttatgcagct gcctaccagc ttcaccctca tgcctggtgg ggcagtggcc cagcaggtcc
     1201 ctgtgcaggc cattcatgtg caccaggccc cacagcaagc gtctccctct cgtgacagca
     1261 gcacagacct cacgcagacc tcctccagcg ggacagtgac gttgcccgcc accatcatga
     1321 cgtcgtctgt acccacaact gtgggtggcc acatgatgta ccctagtccc catgcagtga
     1381 tgtatgcccc cacctcaggc ctggctgatg gcagcctcac cgtgctcaat gccttctctc
     1441 aggcaccatc caccatgcag gtgtcccaca gccaggtcca ggagccaggt ggtgtccctc
     1501 aggtgttcct gacagcaccg tctgggaccg tgcagatccc tgtctctgca gttcagcttc
     1561 accagatggc tgtgataggg cagcaagctg ggagcagcag caacctcacc gagctacagg
     1621 tggtgaacct ggatgccacc cacagcacca agagtgaaaa gggcgaattc gacccagctt
     1681 tcttgtacaa agttggcatt ataaaaaata attgctcatc aatttgttgc aacgaacagg
     1741 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     1801 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1861 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     1921 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     1981 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     2041 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     2101 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     2161 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     2221 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     2281 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     2341 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     2401 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     2461 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     2521 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     2581 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     2641 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     2701 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     2761 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     2821 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     2881 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     2941 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     3001 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     3061 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3361 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3421 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3481 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3541 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3601 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3661 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3721 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3781 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3841 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3901 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3961 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     4021 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     4081 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     4141 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     4201 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     4261 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     4321 cacgacgtt
//
