LOCUS       sequence-442832-        3906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-1 W208L DNA binding mutation overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3906)
  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 3906)
  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_W208L
FEATURES             Location/Qualifiers
     source          1..3906
                     /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(1217..1316)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1333..1352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1334..1352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1357..1373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1357..1373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1804..2595
                     /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      2688..3276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3177..3196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3430..3447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3606..3633)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3725..3811)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3860..3882
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3874..3891
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3875..3891
                     /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 atcttgttcc agaaccaccg ctacaaaatg aagcgccagg ccaaggacaa ggcggcgcag
      781 cagcaactgc agcaggacag cggcggcggc gggggcggcg ggggcaccgg gtgcccgcag
      841 cagcaacagg ctcagcagca gtcgccgcga cgcgtggcgg tgccggtcct ggtgaaagac
      901 ggcaaaccgt gccaggcggg tgcccccgcg ccgggcgccg ccagcctaca aggccacgcg
      961 cagcagcagg cgcagcacca ggcgcaggcc gcgcaggcgg cggcagcggc catctccgtg
     1021 ggcagcggtg gcgccggcct tggcgcacac ccgggccacc agccaggcag cgcaggccag
     1081 tctccggacc tggcgcacca cgccgccagc cccgcggcgc tgcagggcca ggtatccagc
     1141 ctgtcccacc tgaactcctc gggctcggac tacggcacca tgtcctgctc caccttgcta
     1201 tacggtcgga cctggttgcc aactttcttg tacaaagttg gcattataag aaagcattgc
     1261 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1321 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1381 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatgc
     1441 ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg
     1501 tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt
     1561 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
     1621 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
     1681 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
     1741 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
     1801 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1861 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1921 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1981 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     2041 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     2101 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     2161 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     2221 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     2281 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     2341 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     2401 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     2461 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     2521 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     2581 gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg
     2641 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2701 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2761 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2821 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2881 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2941 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3001 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3061 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3301 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3361 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3421 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3481 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3541 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3601 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     3661 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     3721 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     3781 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     3841 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     3901 gggccc
//
