LOCUS       sequence-182702-        4275 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gateway entry TOPO TA cloning vector for ETV6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4275)
  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 4275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ETV6
FEATURES             Location/Qualifiers
     source          1..4275
                     /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(1618..1717)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1734..1753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1735..1753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1758..1774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1758..1774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2205..2996
                     /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      3089..3677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3578..3597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3831..3848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4007..4034)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4126..4212)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4261..4275,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4275,1..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 tgtctgagac tcctgctcag tctagcatta
      181 agcaggaacg aatttcatac acgcccccag agagtccagt ggcaagccac cgttcctcga
      241 ctccgcttca tgttcacaca gtgcctcgag cgctcaggat ggaggaagac tcgatccacc
      301 tgccaacaca cctgcgtttg cagccgattt actggagcag agatgacgta gcccagtggc
      361 tcaagtgggc agaaaatgag ttttctttaa ggcccattga gagcaacaag ttcgaaatga
      421 atggcaaggc cctcctgctg ctgaccaaag aggatttccg ctaccgatct cctcattcag
      481 gcgacgtgct ctatgaactc cttcagcata tcctgaagca gaggaaatct cgaatgctct
      541 tctcaccatt cttcccccct ggggactcta tccacaccaa gccagaggtc ctcctgcatc
      601 agaaccatga cgaagataac tgtgtccaga ggacacccag gacgcccgcg gagagcgtgc
      661 accacaaccc tcccaccatc gaactcttac atcgccctag gtcacccatc accacaaacc
      721 acaggccttc tcctgacccc gaacagcagc ggccccagcg gtccccccta gacaacatga
      781 gccgccgcct ctcgccagtg gagaaagccc aggggcccag gctacagcag gagaacaacc
      841 accaggaaac gtaccccctg tcagtgtctc ctgtcgagaa taatcactgc ctgccctcaa
      901 gcccctggca ggagagcact cgagtgatcc agctgatgcc cagccccatc atgcaccctt
      961 tgatcctgaa cccccggcac tcgcactcgg tggacttcaa acagtcccgg cactccgagg
     1021 atgggatgaa tcgggaaggg aagcccatca acctgtctca tcgggaggac ctggcttact
     1081 tgaaccacat catggtctct atgtccccac cggaagagca cgccatgccc attgggagaa
     1141 tagcagactg tagactgctt tgggattatg tctatcagtt gctgtctgac agccggtacg
     1201 aaaacttcat ccgatgggag gacaaagaat ccaaaatatt ccggatagtg gatcccaacg
     1261 gactggctcg actctgggga aaccataaga acagaacaaa catgacctat gagaaaatgt
     1321 ccagagccct gcgccactac tacaaactaa acattatcag gaaggagccc ggacaaaggc
     1381 ttttgttcag gttcatgaaa accccagatg agatcatgag tggccggaca gaccgtctag
     1441 aacacctcga gtctcaagtg ctggatgaac aaacgtacca agaggatgaa cctaccatag
     1501 cctcaccggt gggctggcca agaggaaacc tgcccacggg gaccgcagga ggcgtgatgg
     1561 aagcaggcga gctaggggtg gctgtaaagg aagagacccg ggaaaagggc gaattcgacc
     1621 cagctttctt gtacaaagtt ggcattataa aaaataattg ctcatcaatt tgttgcaacg
     1681 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1741 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1801 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     1861 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     1921 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     1981 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     2041 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     2101 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     2161 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     2221 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     2281 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     2341 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     2401 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     2461 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2521 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2581 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     2641 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     2701 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     2761 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     2821 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     2881 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     2941 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     3001 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3841 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3901 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3961 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     4021 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     4081 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4141 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4201 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     4261 cccagtcacg acgtt
//
