LOCUS       sequence-442839-        2988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-1 N-terminal and C-terminal truncation 
            (homeodomain only) mutation overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2988)
  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 2988)
  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_HD_160-223AA
FEATURES             Location/Qualifiers
     source          1..2988
                     /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(299..398)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(415..434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(416..434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(439..455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(439..455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             886..1677
                     /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      1770..2358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2259..2278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2512..2529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2688..2715)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2807..2893)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2942..2964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2956..2973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2957..2973
                     /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 ggacaagttg
      301 ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
      361 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
      421 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
      481 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag
      541 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
      601 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
      661 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
      721 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
      781 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
      841 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
      901 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
      961 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     1021 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     1081 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     1141 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     1201 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     1261 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     1321 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     1381 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     1441 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     1501 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     1561 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     1621 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     1681 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     1741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2461 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2521 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2641 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     2701 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     2761 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     2821 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     2881 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     2941 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
