LOCUS       sequence-182583-        4155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry TOPO TA cloning vector for TBX20a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4155)
  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 4155)
  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-TBX20a
FEATURES             Location/Qualifiers
     source          1..4155
                     /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(1498..1597)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1614..1633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1615..1633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1638..1654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1638..1654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2085..2876
                     /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      2969..3557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3458..3477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3711..3728
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3887..3914)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4006..4092)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4141..4155,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4155,1..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 tggagttcac ggcgtcgccc aagccccagc
      181 tctcctctcg agccaatgcc ttctccatcg ccgcgcttat gtccagcggc ggccccaagg
      241 agaaggaggc agcagagaac accatcaaac ccctggaaca atttgtggag aaatcatcat
      301 gtgcccagcc actgggtgag ctgacgagtc tggatgctca cgcggagttt ggcggcgggg
      361 gcggcagccc atcctcatcc tctctgtgca cagagccact gatacccacc acccccatca
      421 tccccagcga agagatggct aaaatcgcct gcagcctgga aacgaaggag ctctgggaca
      481 aattccatga actgggcacg gagatgataa tcaccaagtc tggcaggagg atgttcccca
      541 ccatccgcgt gtcattttct ggagtggatc ctgagtccaa gtatatagtc ctgatggaca
      601 tcgtcccggt ggacaacaag agataccgct atgcctacca ccggtcatcc tggctggtgg
      661 ctggcaaagc tgatcccccg ctgccagcca ggctctacgt gcacccagac tcccccttta
      721 ctggcgagca gctcctcaaa cagatggtgt cttttgaaaa ggtgaagctc accaacaatg
      781 aactggatca acacggccat ataattttga attcaatgca taagtaccag ccacgggtgc
      841 acatcataaa gaagaaagac cacacggcct ccttgctcaa tctgaagtca gaagaattca
      901 ggacgttcat ctttccagag acagttttca cagcagtcac agcctaccag aaccaactga
      961 taaccaagct gaaaatagac agcaatccgt ttgccaaagg attccgggac tcctccaggc
     1021 tcactgacat tgagagggag agtgtggaga gcctgatcca gaagcattcc tatgcccggt
     1081 cacccatccg cacctatggg gaagaggatg ttctggggga ggagagtcaa acaactcaga
     1141 gtcgaggatc agcctttaca acatctgaca atttgtctct cagttcctgg gtatcgtcat
     1201 cttccagttt tcctggattt cagcatccac agcccctgac tgctcttggt accagtacag
     1261 catccatagc gacaccgatt cctcacccta tacagggttc tctgccacca tatagccgcc
     1321 tgggaatgcc tctgacccca tctgcaatag ccagctccat gcagggaagt ggtcccacgt
     1381 tcccttcatt ccacatgcct agataccatc actacttcca gcaggggccc tacgctgcca
     1441 tccaaggact tcgccactcc tccgctgtga tgacaccatt tgtaaagggc gaattcgacc
     1501 cagctttctt gtacaaagtt ggcattataa aaaataattg ctcatcaatt tgttgcaacg
     1561 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1621 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1681 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     1741 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     1801 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     1861 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     1921 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     1981 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     2041 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     2101 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     2161 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     2221 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     2281 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     2341 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2401 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2461 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     2521 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     2581 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     2641 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     2701 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     2761 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     2821 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     2881 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     2941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3661 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3721 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3781 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3841 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     3901 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     3961 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4021 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4081 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     4141 cccagtcacg acgtt
//
