LOCUS       sequence-442838-        3765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway cloning of NKX2-5 overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3765)
  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 3765)
  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_NKX2-5
FEATURES             Location/Qualifiers
     source          1..3765
                     /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(1076..1175)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1192..1211)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1193..1211)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1216..1232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1216..1232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1663..2454
                     /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      2547..3135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3036..3055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3289..3306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3465..3492)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3584..3670)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3719..3741
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3733..3750
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3734..3750
                     /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 ccatgttccc cagccctgct
      121 ctcacgccca cgcccttctc agtcaaagac atcctaaacc tggaacagca gcagcgcagc
      181 ctggctgccg ccggagagct ctctgcccgc ctggaggcga ccctggcgcc ctcctcctgc
      241 atgctggccg ccttcaagcc agaggcctac gctgggcccg aggcggctgc gccgggcctc
      301 ccagagctgc gcgcagagct gggccgcgcg ccttcaccgg ccaagtgtgc gtctgccttt
      361 cccgccgccc ccgccttcta tccacgtgcc tacagcgacc ccgacccagc caaggaccct
      421 agagccgaaa agaaagagct gtgcgcgctg cagaaggcgg tggagctgga gaagacagag
      481 gcggacaacg cggagcggcc ccgggcgcga cggcggagga agccgcgcgt gctcttctcg
      541 caggcgcagg tctatgagct ggagcggcgc ttcaagcagc agcggtacct gtcggccccc
      601 gaacgcgacc agctggccag cgtgctgaaa ctcacgtcca cgcaggtcaa gatctggttc
      661 cagaaccggc gctacaagtg caagcggcag cggcaggacc agactctgga gctggtgggg
      721 ctgcccccgc cgccgccgcc gcctgcccgc aggatcgcgg tgccagtgct ggtgcgcgat
      781 ggcaagccat gcctagggga ctcggcgccc tacgcgcctg cctacggcgt gggcctcaat
      841 ccctacggtt ataacgccta ccccgcctat ccgggttacg gcggcgcggc ctgcagccct
      901 ggctacagct gcactgccgc ttaccccgcc gggccttccc cagcgcagcc ggccactgcc
      961 gccgccaaca acaacttcgt gaacttcggc gtcggggact tgaatgcggt tcagagcccc
     1021 gggattccgc agagcaactc gggagtgtcc acgctgcatg gtatccgagc ctggttgcca
     1081 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     1141 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     1201 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     1261 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     1321 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     1381 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     1441 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     1501 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     1561 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     1621 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     1681 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     1741 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     1801 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     1861 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     1921 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     1981 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     2041 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     2101 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     2161 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     2221 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     2281 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     2341 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     2401 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     2461 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3241 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3301 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3361 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3421 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3481 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3541 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3601 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3661 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     3721 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
