LOCUS       sequence-117049-        7727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To create a dox-on inducible expression of human wt-TTF-1 cDNA in 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  AUTHORS   Rice SJ, Lai SC, Wood LW, Helsley KR, Runkle EA, Winslow MM, Mu D
  TITLE     MicroRNA-33a mediates the regulation of high mobility group AT-hook 
            2 gene (HMGA2) by thyroid transcription factor 1 (TTF-1/NKX2-1).
  JOURNAL   J Biol Chem. 2013 Jun 7;288(23):16348-60. doi: 
            10.1074/jbc.M113.474643. Epub 2013 Apr 26.
  PUBMED    23625920
REFERENCE   2  (bases 1 to 7727)
  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"; doi: 
            "10.1074/jbc.M113.474643"; journalName: "J Biol Chem"; date: 
            "2013-06-7- 7"; volume: "288"; issue: "23"; pages: "16348-60"
COMMENT     Sequence Label: Puro-TTL-wt-TTF-1
FEATURES             Location/Qualifiers
     source          1..7727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             514..689
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    752..951
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1152..1568
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1498..1520
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1536..1552
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1600..1914
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1608..1626
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1644..1662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1679..1697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1715..1733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1751..1769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1786..1804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1822..1840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1887..1911
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3113..3612
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3148..3171)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3545..3564
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3633..4232
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3633..3652)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4129..4149
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4433..4858
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4961..4983)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5119..5139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5124..5453
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5304..5439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5475..5494
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5575..5592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5746..6334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5826..5845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6505..7296,7297..7365))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7128..7147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7366..7470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7538..7556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7656..7727,1..309)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact
      541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
      601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
      721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
      781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
      841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1561 taggcgccgg aattgaagat ctgggggatc gataccgtcg agtttactcc ctatcagtga
     1621 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc
     1681 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
     1741 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata
     1801 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt
     1861 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     1921 attaattcga gctcggtacc cggggatcct ccggaactta gcagtctcga gaccatgtcg
     1981 atgagtccaa agcacacgac tccgttctca gtgtctgaca tcttgagtcc cctggaggaa
     2041 agctacaaga aagtgggcat ggagggcggc ggcctcgggg ctccgctggc ggcgtacagg
     2101 cagggccagg cggcaccgcc aacagcggcc atgcagcagc acgccgtggg gcaccacggc
     2161 gccgtcaccg ccgcctacca catgacggcg gcgggggtgc cccagctctc gcactccgcc
     2221 gtggggggct actgcaacgg caacctgggc aacatgagcg agctgccgcc gtaccaggac
     2281 accatgagga acagcgcctc tggccccgga tggtacggcg ccaacccaga cccgcgcttc
     2341 cccgccatct cccgcttcat gggcccggcg agcggcatga acatgagcgg catgggcggc
     2401 ctgggctcgc tgggggacgt gagcaagaac atggccccgc tgccaagcgc gccgcgcagg
     2461 aagcgccggg tgctcttctc gcaggcgcag gtgtacgagc tggagcgacg cttcaagcaa
     2521 cagaagtacc tgtcggcgcc ggagcgcgag cacctggcca gcatgatcca cctgacgccc
     2581 acgcaggtca agatctggtt ccagaaccac cgctacaaaa tgaagcgcca ggccaaggac
     2641 aaggcggcgc agcagcaact gcagcaggac agcggcggcg gcgggggcgg cgggggcacc
     2701 gggtgcccgc agcagcaaca ggctcagcag cagtcgccgc gacgcgtggc ggtgccggtc
     2761 ctggtgaaag acggcaaacc gtgccaggcg ggtgcccccg cgccgggcgc cgccagccta
     2821 caaggccacg cgcagcagca ggcgcagcac caggcgcagg ccgcgcaggc ggcggcagcg
     2881 gccatctccg tgggcagcgg tggcgccggc cttggcgcac acccgggcca ccagccaggc
     2941 agcgcaggcc agtctccgga cctggcgcac cacgccgcca gccccgcggc gctgcagggc
     3001 caggtatcca gcctgtccca cctgaactcc tcgggctcgg actacggcac catgtcctgc
     3061 tccaccttgc tatacggtcg gacctggtga ctcgaggtta acgaattcta ccgggtaggg
     3121 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     3181 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     3241 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     3301 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     3361 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     3421 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     3481 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     3541 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     3601 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     3661 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     3721 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     3781 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3841 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     3901 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     3961 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4021 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4081 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4141 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4201 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     4261 ccgaaaggag cgcacgaccc catgcatcga atcgcgtcct gcaggccgat accgtcgaga
     4321 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt
     4381 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     4441 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     4501 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagggtcga
     4561 ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     4621 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     4681 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     4741 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     4801 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
     4861 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
     4921 aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4981 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5041 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     5101 gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5221 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5281 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5341 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5401 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
     5461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     5581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6181 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     6841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     7141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7561 tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata atatgtacat
     7621 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
     7681 atagtaatca attacggggt cattagttca tagcccatat atggagt
//
