LOCUS       sequence-437812-        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"
     rep_origin      390..978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     879..898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1132..1149
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1308..1335)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1427..1513)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1562..1584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1576..1593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1577..1593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1609..1708
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2773..2872)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2889..2908)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2890..2908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2913..2929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3360..3854,1..297)
                     /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"
ORIGIN
        1 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
       61 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
      121 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
      181 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
      241 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
      301 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
      361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
      421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1081 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1141 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1201 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1261 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     1321 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     1381 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     1441 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     1501 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     1561 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt
     1621 ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata
     1681 atgccaactt tgtacaaaaa agttggcaca tgtcgttgag cccaaagcac acgacgccct
     1741 tctccgtgtc cgacatcctg agccccatcg aggagaccta caagaagttc agcggcgcca
     1801 tggacggcgc gccacccggc ctgggggcgc ccctgggggc cgcggccgcc taccgcgcgc
     1861 cgccacctgg gccctcctcg caggcggcga ccgtggcggg catgcagcct tctcacgcca
     1921 tggcgggtca caacgcggcg gccgcggcgg cggcggcagc agctgcggcg gcggcggccg
     1981 ccacctacca catgccgccc ggcgtctcgc agttcccgca cggcgccatg ggcagctact
     2041 gcaacggcgg cctgggcaac atgggcgagc tgcccgccta cacggacggc atgcggggcg
     2101 gcgcggccac cggctggtac ggcgccaacc cggacccacg ctactcgtca atctccaggt
     2161 tcatggggcc gtcggcgggc gtgaatgtgg ccggcatggg gtcgctgacc ggcatcgcgg
     2221 acgccgccaa gtcgctgggc ccgctgcacg cggcggcggc ggcagccgct ccgcgaagga
     2281 agcgccgcgt gctcttctcg caggcgcagg tctacgagct ggagcggcgc ttcaagcagc
     2341 agaagtacct gtcggcgccc gagcgcgagc acctggccag catgatccac ctgacgccca
     2401 cgcaggtcaa gatctggttc cagaaccacc ggtacaagat gaaacggcag gccaaggaca
     2461 aggcggcgca gcagctgcag caggagggcg gcctgggccc gccgccgcct ccgccgccgt
     2521 ccccgcgccg cgtggcggtg cctgtgctgg tcaaggacgg caagccgtgc cagaacggcg
     2581 ccagcacgcc cacccccggc caggccggtc cgcagccgcc ggccccgacg ccagcacctg
     2641 agctggagga gctgtcgccc agcccacccg cgctgcacgg cccggggggc ggcctggcgg
     2701 ccctggacgc ggccgccggg gagtacagcg gcggcgtcct gggcgccaac ctgctctatg
     2761 gcaggacgtg gttgccaact ttcttgtaca aagttggcat tataagaaag cattgcttat
     2821 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     2881 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2941 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     3001 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     3061 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     3121 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     3181 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     3241 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     3301 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     3361 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     3421 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     3481 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     3541 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     3601 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     3661 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     3721 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     3781 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     3841 atctatttga ggcg
//
