LOCUS       sequence-442834-        3996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-1 long isoform overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3996)
  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 3996)
  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_Long
FEATURES             Location/Qualifiers
     source          1..3996
                     /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(1307..1406)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1423..1442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1424..1442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1447..1463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1447..1463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1894..2685
                     /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      2778..3366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3267..3286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3520..3537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3696..3723)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3815..3901)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3950..3972
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3964..3981
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3965..3981
                     /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 ccatgtggtc cggaggcagt
      121 gggaaggcgc ggggctggga ggccgcggcg ggagggagga gcagccccgg caggctcagc
      181 cgccgccgaa tcatgtcgat gagtccaaag cacacgactc cgttctcagt gtctgacatc
      241 ttgagtcccc tggaggaaag ctacaagaaa gtgggcatgg agggcggcgg cctcggggct
      301 ccgctggcgg cgtacaggca gggccaggcg gcaccgccaa cagcggccat gcagcagcac
      361 gccgtggggc accacggcgc cgtcaccgcc gcctaccaca tgacggcggc gggggtgccc
      421 cagctctcgc actccgccgt ggggggctac tgcaacggca acctgggcaa catgagcgag
      481 ctgccgccgt accaggacac catgaggaac agcgcctctg gccccggatg gtacggcgcc
      541 aacccagacc cgcgcttccc cgccatctcc cgcttcatgg gcccggcgag cggcatgaac
      601 atgagcggca tgggcggcct gggctcgctg ggggacgtga gcaagaacat ggccccgctg
      661 ccaagcgcgc cgcgcaggaa gcgccgggtg ctcttctcgc aggcgcaggt gtacgagctg
      721 gagcgacgct tcaagcaaca gaagtacctg tcggcgccgg agcgcgagca cctggccagc
      781 atgatccacc tgacgcccac gcaggtcaag atctggttcc agaaccaccg ctacaaaatg
      841 aagcgccagg ccaaggacaa ggcggcgcag cagcaactgc agcaggacag cggcggcggc
      901 gggggcggcg ggggcaccgg gtgcccgcag cagcaacagg ctcagcagca gtcgccgcga
      961 cgcgtggcgg tgccggtcct ggtgaaagac ggcaaaccgt gccaggcggg tgcccccgcg
     1021 ccgggcgccg ccagcctaca aggccacgcg cagcagcagg cgcagcacca ggcgcaggcc
     1081 gcgcaggcgg cggcagcggc catctccgtg ggcagcggtg gcgccggcct tggcgcacac
     1141 ccgggccacc agccaggcag cgcaggccag tctccggacc tggcgcacca cgccgccagc
     1201 cccgcggcgc tgcagggcca ggtatccagc ctgtcccacc tgaactcctc gggctcggac
     1261 tacggcacca tgtcctgctc caccttgcta tacggtcgga cctggttgcc aactttcttg
     1321 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1381 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1441 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1501 attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc
     1561 tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
     1621 cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc
     1681 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     1741 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     1801 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     1861 cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta
     1921 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     1981 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     2041 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     2101 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     2161 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     2221 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     2281 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     2341 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     2401 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     2461 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     2521 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     2581 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     2641 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca
     2701 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3421 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3481 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3541 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3601 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3661 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3721 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3781 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3841 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3901 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3961 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
