LOCUS       sequence-237539-        8205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible mouse ID1 from 806 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  AUTHORS   Huang YH, Hu J, Chen F, Lecomte N, Basnet H, David CJ, Witkin MD, 
            Allen PJ, Leach SD, Hollmann TJ, Iacobuzio-Donahue CA, Massague J
  TITLE     ID1 mediates escape from TGF-beta tumor suppression in pancreatic 
            cancer.
  JOURNAL   Cancer Discov. 2019 Oct 3. pii: 2159-8290.CD-19-0529. doi: 
            10.1158/2159-8290.CD-19-0529.
  PUBMED    31582374
REFERENCE   2  (bases 1 to 8205)
  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: "Cancer 
            Discov. 2019 Oct 3. pii: 2159-8290.CD-19-0529. doi: 
            10.1158/2159-8290.CD-19-0529."
COMMENT     Sequence Label: ID1-pLVXT
FEATURES             Location/Qualifiers
     source          1..8205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..322
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    16..34
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    52..70
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    87..105
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    123..141
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    159..177
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    194..212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    230..248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     295..319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        793..1292
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(828..851)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1225..1244
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1313..1912
                     /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(1313..1332)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1809..1829
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1929..2517
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1982..2002)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2400..2411)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2724..3357
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3485..3501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3485..3501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3498..3520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3509..3525
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3533..3539,3540..3557,3558..3563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3578..3599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3716..3733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3887..4475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3967..3986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4646..5437,5438..5506))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5269..5288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5507..5611)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5659..5793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5696..5715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5750..5769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6008..6641
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6688..6813
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7310..7543
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7728..7772
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8035..8150
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 ttcactcgag tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca
       61 gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      121 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      181 tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      241 gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa gcagagctcg
      301 tttagtgaac cgtcagatcg cctggagaag gatccatgaa ggtcgccagt ggcagtgccg
      361 cagccgctgc aggccctagc tgttcgctga aggcgggcag gacagcgggc gaggtggtac
      421 ttggtctgtc ggagcaaagc gtggccatct cgcgctgcgc tgggacgcgc ctgcccgcct
      481 tgctggacga gcagcaggtg aacgtcctgc tctacgacat gaacggctgc tactcacgcc
      541 tcaaggagct ggtgcccacc ctgccccaga accgcaaagt gagcaaggtg gagatcctgc
      601 agcatgtaat cgactacatc agggacctgc agctggagct gaactcggag tctgaagtcg
      661 ggaccaccgg aggccgggga ctgcctgtcc gcgccccgct cagcaccctg aacggcgaga
      721 tcagtgcctt ggcggccgag gcggcatgtg ttccagccga cgatcgcatc ttgtgtcgct
      781 gagaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
      841 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
      901 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
      961 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     1021 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     1081 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     1141 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     1201 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     1261 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga
     1321 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     1381 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     1441 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     1501 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     1561 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     1621 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     1681 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     1741 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     1801 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     1861 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgggcgcg
     1921 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1981 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2041 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2101 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2161 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2221 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2281 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     2341 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2401 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2461 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     2521 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     2581 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     2641 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     2701 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     2761 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     2821 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     2881 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     2941 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     3001 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     3061 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     3121 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     3181 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     3241 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     3301 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     3361 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     3421 gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc ttggcgtaat
     3481 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3541 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3601 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3661 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3721 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3781 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4321 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5521 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5581 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5641 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5701 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5761 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     5821 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
     5881 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgywy twrwymwrtr
     5941 wktmctmyra aytwwktwsw tkkwaargaa attgtatttg ttaaatatgt actacaaact
     6001 tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc
     6061 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga
     6121 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa
     6181 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat
     6241 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg
     6301 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga
     6361 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc
     6421 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca
     6481 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6541 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6601 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     6661 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     6721 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     6781 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     6841 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     6901 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     6961 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     7021 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     7081 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     7141 gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     7201 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     7261 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     7321 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     7381 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     7441 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     7501 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     7561 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     7621 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     7681 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     7741 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     7801 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     7861 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     7921 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     7981 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cgcctttaaa
     8041 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     8101 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     8161 tacagggaca gcagagatcc agtttatcga tgaggccctt tcgtc
//
