LOCUS       sequence-442836-        3432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-1 N-terminal truncation mutation 
            overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3432)
  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 3432)
  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_DelNTerm_160-371AA
FEATURES             Location/Qualifiers
     source          1..3432
                     /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(743..842)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(859..878)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(860..878)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(883..899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(883..899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1330..2121
                     /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      2214..2802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2703..2722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2956..2973
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3132..3159)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3251..3337)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3386..3408
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3400..3417
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3401..3417
                     /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 ccatgccgcg caggaagcgc
      121 cgggtgctct tctcgcaggc gcaggtgtac gagctggagc gacgcttcaa gcaacagaag
      181 tacctgtcgg cgccggagcg cgagcacctg gccagcatga tccacctgac gcccacgcag
      241 gtcaagatct ggttccagaa ccaccgctac aaaatgaagc gccaggccaa ggacaaggcg
      301 gcgcagcagc aactgcagca ggacagcggc ggcggcgggg gcggcggggg caccgggtgc
      361 ccgcagcagc aacaggctca gcagcagtcg ccgcgacgcg tggcggtgcc ggtcctggtg
      421 aaagacggca aaccgtgcca ggcgggtgcc cccgcgccgg gcgccgccag cctacaaggc
      481 cacgcgcagc agcaggcgca gcaccaggcg caggccgcgc aggcggcggc agcggccatc
      541 tccgtgggca gcggtggcgc cggccttggc gcacacccgg gccaccagcc aggcagcgca
      601 ggccagtctc cggacctggc gcaccacgcc gccagccccg cggcgctgca gggccaggta
      661 tccagcctgt cccacctgaa ctcctcgggc tcggactacg gcaccatgtc ctgctccacc
      721 ttgctatacg gtcggacctg gttgccaact ttcttgtaca aagttggcat tataagaaag
      781 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      841 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
      901 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
      961 tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt
     1021 gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt
     1081 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     1141 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     1201 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     1261 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     1321 ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
     1381 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     1441 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
     1501 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     1561 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     1621 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     1681 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     1741 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     1801 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     1861 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     1921 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     1981 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     2041 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
     2101 accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt
     2161 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2221 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2281 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2341 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2401 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2461 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2521 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2581 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2641 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2701 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2761 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2821 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2881 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2941 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3001 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3061 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     3121 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     3181 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     3241 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     3301 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     3361 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     3421 aagctcgggc cc
//
