LOCUS       7TFP_CDH1_report        9174 bp DNA     circular SYN 13-MAY-2021
DEFINITION  lentiviral luciferase reporter containing CDH1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    7TFP CDH1 reporter.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9174)
  AUTHORS   Pattabiraman DR, Bierie B, Kober KI, Thiru P, Krall JA, Zill C, 
            Reinhardt F, Tam WL, Weinberg RA
  TITLE     Activation of PKA leads to mesenchymal-to-epithelial transition and 
            loss of tumor-initiating ability.
  JOURNAL   Science. 2016 Mar 4;351(6277):aad3680. doi: 10.1126/science.aad3680.
  PUBMED    26941323
REFERENCE   2  (bases 1 to 9174)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9174)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2016-03-4"; pages: "
            10.1126/science.aad3680"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        325..356
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             418..2067
                     /label=luciferase
                     /note="firefly luciferase"
     promoter        2094..2423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2433..3029
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    3099..3687
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3774..4007
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4079..4213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4240..4375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4396..4414)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4424..4440)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4582..5037
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5063..5167
                     /label=AmpR promoter
     CDS             5168..6025
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6199..6787
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6941..6958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7075..7096
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7111..7141
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7149..7165
                     /label=lac operator
                     /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)."
     primer_bind     7173..7189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7210..7228
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7256..7482
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7483..7663
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7710..7835
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8328..8561
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8746..8790
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             8939..8980
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    9057..9174
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 atcgatcacg agactagcct cctagcccgg gctcgagcag ctgaagcttg catgcctgca
       61 ggcgtccgcg ctgctgattg gctgtggccg gcaggtgaac cctcagccaa tcagcggtac
      121 ggggggcggt gcctccgggg ctcacctggc tgcagccacg caccccctct cagtggcgtc
      181 ggaactgcaa agcacctgtg agcttgcgga agtcagttca gactccagcc cgctccagcc
      241 cggcccgacc cgaccgcacc cggcgcctgc cctcgctcgg cgtccccggc agccaccgct
      301 agcccgggct cgagatctag actctagagg gtatataatg gaagctcgaa ttccagcttg
      361 gcattccggt actgttggta aaaagcttgg cattccggta ctgttggtaa agccaccatg
      421 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
      481 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
      541 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
      601 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
      661 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
      721 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
      781 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
      841 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
      901 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
      961 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     1021 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     1081 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     1141 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     1201 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     1261 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     1321 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     1381 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     1441 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     1501 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     1561 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     1621 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     1681 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     1741 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     1801 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     1861 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     1921 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     1981 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     2041 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc gaccctgtgg aatgtgtgtc
     2101 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2161 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2221 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2281 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2341 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2401 ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc
     2461 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     2521 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     2581 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     2641 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     2701 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     2761 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     2821 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     2881 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     2941 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3001 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     3061 ccgaaaggag cgcacgaccc catgcatcgc cagtcgacaa tcaacctctg gattacaaaa
     3121 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3181 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3241 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3301 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3361 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3421 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3481 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     3541 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3601 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3661 ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacctt taagaccaat
     3721 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     3781 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     3841 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     3901 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3961 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     4021 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4201 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     4261 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4321 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     4381 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     4441 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4501 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4561 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4621 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4681 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4741 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4801 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4861 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4921 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4981 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5701 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6121 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6181 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6241 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6301 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6361 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6421 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6481 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6541 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6601 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6661 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6721 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6781 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6841 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6901 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6961 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     7021 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     7081 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     7141 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     7201 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
     7261 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
     7321 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
     7381 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
     7441 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
     7501 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     7561 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     7621 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     7681 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     7741 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     7801 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     7861 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     7921 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     7981 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     8041 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     8101 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     8161 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     8221 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     8281 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     8341 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     8401 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     8461 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     8521 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     8581 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     8641 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     8701 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     8761 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     8821 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     8881 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     8941 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     9001 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt
     9061 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     9121 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa tttt
//
