LOCUS       V016414                 8403 bp    DNA     circular SYN 26-MAY-2020
DEFINITION  Exported.
ACCESSION   V016414
VERSION     V016414
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8403)
  AUTHORS   Han X, Yang J, Zeng F, Weng J, Zhang Y, Peng Q, Shen L, Ding S, Liu
            K, Gao Y.
  TITLE     Programmable Synthetic Protein Circuits for the Identification and
            Suppression of Hepatocellular Carcinoma
  JOURNAL   Mol Ther Oncolytics 17, 70-82 (2020)
   PUBMED   32322664
REFERENCE   2  (bases 1 to 8403)
  AUTHORS   Liu K, Yang J, Ding S.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2020) Second Department of Hepatobiliary Surgery,
            Zhujiang Hospital, No. 253, Industrial Avenue, Haizhu District,
            Guangzhou, Guangdong 510245, China
REFERENCE   3  (bases 1 to 8403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther
            Oncolytics 17, 70-82 (2020)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-JAN-2020) Second Department of Hepatobiliary Surgery, Zhujiang
            Hospital, No. 253, Industrial Avenue, Haizhu District, Guangzhou,
            Guangdong 510245, China"
FEATURES             Location/Qualifiers
     source          1..8403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             884..1204
                     /label="DD"
                     /note="destabilization domain that can be stabilized by
                     Shield1 in the ProteoTuner(TM) system"
     CDS             1322..1342
                     /label="TVMV site"
                     /note="tobacco vein mottling virus (TVMV) NIa protease
                     recognition and cleavage site"
     CDS             1355..1741
                     /label="MS2-N55K"
                     /note="bacteriophage MS2 coat protein"
     CDS             1754..2608
                     /gene="CNOT7"
                     /label="CCR4-NOT transcription complex subunit 7"
                     /note="CCR4-NOT transcription complex subunit 7 from Homo
                     sapiens. Accession#: Q9UIV1"
     CDS             2627..2686
                     /label="E2A"
                     /note="2A peptide from equine rhinitis A virus polyprotein"
     CDS             2699..4348
                     /label="luciferase"
                     /note="firefly luciferase"
     polyA_signal    4399..4623
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4669..5097
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5111..5440
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5507..5902
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    6079..6212
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6265)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6273..6289)
                     /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)."
     promoter        complement(6297..6327)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6342..6363)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6651..7236)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7410..8267)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8268..8372)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctaggaatt cgcggccgcg ccaccatggg cagatctatg ggtggtgtgc aggttgaaac
      901 tatatccccg ggtgatggtc gaacattccc caaacgaggg cagacatgcg tcgtgcacta
      961 tactggaatg ctggaggatg gaaagaaagt ggacagtagc cgagatcgta acaaaccatt
     1021 caaattcatg ctggggaaac aggaggttat ccggggatgg gaggagggtg tggcccagat
     1081 gtctgtgggc cagcgggcta aactgaccat aagccccgac tacgcctacg gagccacagg
     1141 gcacccaggt attatccccc cacacgctac cctggtcttt gacgttgagc ttctcaagcc
     1201 tgagggaggc ggttccgaac aattggagaa gaagcttcaa gctctggaaa aaaaactcgc
     1261 ccagctggag tggaagaacc aggccttaga gaagaaacta gcccaaggag gctctggatc
     1321 tgagacggtt agattccaga gtggctcagg atctgcctca aactttacac agtttgtatt
     1381 agtggacaac ggcggcaccg gcgatgtgac cgtcgctcca tcgaattttg caaacggtgt
     1441 ggccgagtgg attagctcga actccagaag ccaggcctat aaagtaacat gctccgtgag
     1501 gcaaagcagc gctcagaaac ggaagtacac tataaaggtg gaggtgccga aggtggcaac
     1561 ccagaccgtg ggcggggtgg aactgcctgt tgctgcctgg cgatcctacc taaatatgga
     1621 gctgaccatt cccatattcg ctaccaattc cgactgtgag ctaatagtca aagccatgca
     1681 ggggctactg aaagatggga atccgatccc ttccgctata gccgctaatt ccgggatcta
     1741 tggatctatc gctatgccgg ctgccaccgt tgaccactca caaaggatct gcgaagtgtg
     1801 ggcctgtaac cttgatgagg agatgaagaa gatcagacag gtcattagga agtacaacta
     1861 tgttgctatg gacacagagt tcccaggagt ggtcgcaagg cctattggcg agttccggag
     1921 caatgcagat tatcagtatc agttgttgag gtgtaacgtg gatctcttaa aaatcattca
     1981 gctcggactt acattcatga acgaacaggg cgaatatccc cccggcactt ctacatggca
     2041 gttcaatttc aagtttaatc tgacagagga catgtatgcc caggattcga tcgagctgct
     2101 cacaactagc ggaatccagt ttaagaagca cgaggaagaa ggcatcgaaa cccagtactt
     2161 tgcagagctt ttgatgacgt ctggggtggt gctctgtgag ggcgtcaagt ggctcagctt
     2221 tcatagtggt tatgattttg gttatctcat caagatcctg actaacagta atctcccaga
     2281 agaggagcta gactttttcg agatactccg gctcttcttt cccgttatat atgacgtgaa
     2341 atatctcatg aaaagctgta agaacctcaa aggagggctt caggaagtgg cggagcagct
     2401 ggagcttgaa agaattggac cacagcatca ggccgggtct gactcactgc tcaccgggat
     2461 ggccttcttc aaaatgcgcg agatgttttt cgaagatcac attgacgatg ccaagtactg
     2521 cgggcatctg tacgggcttg gcagcggaag ctcatatgtg cagaatggaa ccggaaacgc
     2581 ctacgaggaa gaggccaaca agcagtcagt aggatctgga tccgcgcagt gtactaatta
     2641 tgctctcttg aaattggctg gagatgttga gagcaaccca ggtcccatgg gcagatctat
     2701 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatcctctag aggatggaac
     2761 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
     2821 ttttacagat gcacatatcg aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt
     2881 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
     2941 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
     3001 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgaacatttc
     3061 gcagcctacc gtagtgtttg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
     3121 aaaattacca ataattcaga aaattattat catggattct aaaacggatt accagggatt
     3181 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg agtacgattt
     3241 tgtaccagag tcctttgatc gtgacaaaac aattgcactg ataatgaatt cctctggatc
     3301 tactgggtta cctaagggtg tggcccttcc gcatagaact gcctgcgtca gattctcgca
     3361 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     3421 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     3481 agtcgtctta atgtatagat ttgaagaaga gctgttttta cgatcccttc aggattacaa
     3541 aattcaaagt gcgttgctag taccaaccct attttcattc ttcgccaaaa gcactctgat
     3601 tgacaaatac gatttatcta atttacacga aattgcttct gggggcgcac ctctttcgaa
     3661 agaagtcggg gaagcggttg caaaacgctt ccatcttcca gggatacgac aaggatatgg
     3721 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     3781 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     3841 gctgggcgtt aatcagagag gcgaattatg tgtcagagga cctatgatta tgtccggtta
     3901 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     3961 agacatagct tactgggacg aagacgaaca cttcttcata gttgaccgct tgaagtcttt
     4021 aattaaatac aaaggatatc aggtggcccc cgctgaattg gaatcgatat tgttacaaca
     4081 ccccaacatc ttcgacgcgg gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc
     4141 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     4201 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     4261 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     4321 ggccaagaag ggcggaaagt ccaaattgta accgctcgag aattgttgtt gttgtttaaa
     4381 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4441 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4501 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4561 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4621 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4681 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4741 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4801 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4861 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4921 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4981 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5041 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     5101 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5161 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5281 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5341 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5401 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5461 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatgg ccaagccttt
     5521 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5581 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5641 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5701 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5761 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5821 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5881 ctctggttat gtgtgggagg gctaagcggg actctggggt tcgaaatgac cgaccaagcg
     5941 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6001 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6061 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6181 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6241 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6301 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6361 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6421 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7081 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     7201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8401 gtc
//
