LOCUS       sequence-442841-        3854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-4 overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3854)
  AUTHORS   Pulice JL, Meyerson M
  TITLE     Dosage amplification dictates oncogenic regulation by the NKX2-1 
            lineage factor in lung adenocarcinoma.
  JOURNAL   bioRxiv [Preprint]. 2023 Oct 27:2023.10.26.563996. doi: 
            10.1101/2023.10.26.563996.
  PUBMED    37994369
REFERENCE   2  (bases 1 to 3854)
  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: "bioRxiv 
            [Preprint]. 2023 Oct 27:2023.10.26.563996. doi: 
            10.1101/2023.10.26.563996."
COMMENT     Sequence Label: pDONR223_NKX2-4
FEATURES             Location/Qualifiers
     source          1..3854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1165..1264)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1281..1300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1282..1300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1305..1321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1305..1321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1752..2543
                     /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      2636..3224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3125..3144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3378..3395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3554..3581)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3673..3759)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3808..3830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3822..3839
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3823..3839
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca catgtcgttg agcccaaagc
      121 acacgacgcc cttctccgtg tccgacatcc tgagccccat cgaggagacc tacaagaagt
      181 tcagcggcgc catggacggc gcgccacccg gcctgggggc gcccctgggg gccgcggccg
      241 cctaccgcgc gccgccacct gggccctcct cgcaggcggc gaccgtggcg ggcatgcagc
      301 cttctcacgc catggcgggt cacaacgcgg cggccgcggc ggcggcggca gcagctgcgg
      361 cggcggcggc cgccacctac cacatgccgc ccggcgtctc gcagttcccg cacggcgcca
      421 tgggcagcta ctgcaacggc ggcctgggca acatgggcga gctgcccgcc tacacggacg
      481 gcatgcgggg cggcgcggcc accggctggt acggcgccaa cccggaccca cgctactcgt
      541 caatctccag gttcatgggg ccgtcggcgg gcgtgaatgt ggccggcatg gggtcgctga
      601 ccggcatcgc ggacgccgcc aagtcgctgg gcccgctgca cgcggcggcg gcggcagccg
      661 ctccgcgaag gaagcgccgc gtgctcttct cgcaggcgca ggtctacgag ctggagcggc
      721 gcttcaagca gcagaagtac ctgtcggcgc ccgagcgcga gcacctggcc agcatgatcc
      781 acctgacgcc cacgcaggtc aagatctggt tccagaacca ccggtacaag atgaaacggc
      841 aggccaagga caaggcggcg cagcagctgc agcaggaggg cggcctgggc ccgccgccgc
      901 ctccgccgcc gtccccgcgc cgcgtggcgg tgcctgtgct ggtcaaggac ggcaagccgt
      961 gccagaacgg cgccagcacg cccacccccg gccaggccgg tccgcagccg ccggccccga
     1021 cgccagcacc tgagctggag gagctgtcgc ccagcccacc cgcgctgcac ggcccggggg
     1081 gcggcctggc ggccctggac gcggccgccg gggagtacag cggcggcgtc ctgggcgcca
     1141 acctgctcta tggcaggacg tggttgccaa ctttcttgta caaagttggc attataagaa
     1201 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1261 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1321 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1381 catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     1441 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     1501 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     1561 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     1621 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     1681 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     1741 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     1801 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     1861 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     1921 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     1981 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     2041 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     2101 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     2161 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     2221 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     2281 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     2341 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     2401 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     2461 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     2521 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga
     2581 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3301 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3361 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3421 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3481 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     3541 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     3601 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     3661 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3721 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3781 tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc
     3841 ttaagctcgg gccc
//
