LOCUS       sequence-442833-        3462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-1 C-terminal truncation mutation 
            overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3462)
  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 3462)
  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_NKX2-1_DelCTerm_1-223AA
FEATURES             Location/Qualifiers
     source          1..3462
                     /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(773..872)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(889..908)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(890..908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(913..929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(913..929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1360..2151
                     /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      2244..2832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2733..2752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2986..3003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3162..3189)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3281..3367)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3416..3438
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3430..3447
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3431..3447
                     /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 ccatgtcgat gagtccaaag
      121 cacacgactc cgttctcagt gtctgacatc ttgagtcccc tggaggaaag ctacaagaaa
      181 gtgggcatgg agggcggcgg cctcggggct ccgctggcgg cgtacaggca gggccaggcg
      241 gcaccgccaa cagcggccat gcagcagcac gccgtggggc accacggcgc cgtcaccgcc
      301 gcctaccaca tgacggcggc gggggtgccc cagctctcgc actccgccgt ggggggctac
      361 tgcaacggca acctgggcaa catgagcgag ctgccgccgt accaggacac catgaggaac
      421 agcgcctctg gccccggatg gtacggcgcc aacccagacc cgcgcttccc cgccatctcc
      481 cgcttcatgg gcccggcgag cggcatgaac atgagcggca tgggcggcct gggctcgctg
      541 ggggacgtga gcaagaacat ggccccgctg ccaagcgcgc cgcgcaggaa gcgccgggtg
      601 ctcttctcgc aggcgcaggt gtacgagctg gagcgacgct tcaagcaaca gaagtacctg
      661 tcggcgccgg agcgcgagca cctggccagc atgatccacc tgacgcccac gcaggtcaag
      721 atctggttcc agaaccaccg ctacaaaatg aagcgccagg ccaaggacaa gttgccaact
      781 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
      841 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
      901 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
      961 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
     1021 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1081 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     1141 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     1201 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     1261 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1321 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     1381 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     1441 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     1501 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     1561 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     1621 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     1681 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     1741 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     1801 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     1861 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     1921 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     1981 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2041 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2101 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     2161 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2401 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2461 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2521 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2581 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2641 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2701 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2761 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2821 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2881 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2941 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3001 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3061 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3121 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3181 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3241 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3301 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3361 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3421 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
