LOCUS       V015860                 7695 bp    DNA     circular SYN 26-MAY-2020
DEFINITION  Exported.
ACCESSION   V015860
VERSION     V015860
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7695)
  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 7695)
  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 7695)
  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..7695
                     /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             878..1264
                     /label="MS2-N55K"
                     /note="bacteriophage MS2 coat protein"
     CDS             1271..1291
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             1310..2164
                     /gene="CNOT7"
                     /label="CCR4-NOT transcription complex subunit 7"
                     /note="CCR4-NOT transcription complex subunit 7 from Homo
                     sapiens. Accession#: Q9UIV1"
     CDS             2177..2233
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             2615..2893
                     /label="FRB"
                     /note="FKBP-rapamycin binding domain of human FRAP"
     CDS             2897..2950
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             2951..3271
                     /label="FKBP"
                     /note="human FK506-binding protein FKBP12"
     polyA_signal    3691..3915
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3961..4389
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4403..4732
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4799..5194
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    5371..5504
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5541..5557)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5565..5581)
                     /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(5589..5619)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5634..5655)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5943..6528)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6702..7559)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7560..7664)
                     /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 cagatctgcc tcaaacttta cacagtttgt
      901 attagtggac aacggcggca ccggcgatgt gaccgtcgct ccatcgaatt ttgcaaacgg
      961 tgtggccgag tggattagct cgaactccag aagccaggcc tataaagtaa catgctccgt
     1021 gaggcaaagc agcgctcaga aacggaagta cactataaag gtggaggtgc cgaaggtggc
     1081 aacccagacc gtgggcgggg tggaactgcc tgttgctgcc tggcgatcct acctaaatat
     1141 ggagctgacc attcccatat tcgctaccaa ttccgactgt gagctaatag tcaaagccat
     1201 gcaggggcta ctgaaagatg ggaatccgat cccttccgct atagccgcta attccgggat
     1261 ctatggatct gagaatcttt atttccaatc cggctcagga tctatcgcta tgccggctgc
     1321 caccgttgac cactcacaaa ggatctgcga agtgtgggcc tgtaaccttg atgaggagat
     1381 gaagaagatc agacaggtca ttaggaagta caactatgtt gctatggaca cagagttccc
     1441 aggagtggtc gcaaggccta ttggcgagtt ccggagcaat gcagattatc agtatcagtt
     1501 gttgaggtgt aacgtggatc tcttaaaaat cattcagctc ggacttacat tcatgaacga
     1561 acagggcgaa tatccccccg gcacttctac atggcagttc aatttcaagt ttaatctgac
     1621 agaggacatg tatgcccagg attcgatcga gctgctcaca actagcggaa tccagtttaa
     1681 gaagcacgag gaagaaggca tcgaaaccca gtactttgca gagcttttga tgacgtctgg
     1741 ggtggtgctc tgtgagggcg tcaagtggct cagctttcat agtggttatg attttggtta
     1801 tctcatcaag atcctgacta acagtaatct cccagaagag gagctagact ttttcgagat
     1861 actccggctc ttctttcccg ttatatatga cgtgaaatat ctcatgaaaa gctgtaagaa
     1921 cctcaaagga gggcttcagg aagtggcgga gcagctggag cttgaaagaa ttggaccaca
     1981 gcatcaggcc gggtctgact cactgctcac cgggatggcc ttcttcaaaa tgcgcgagat
     2041 gtttttcgaa gatcacattg acgatgccaa gtactgcggg catctgtacg ggcttggcag
     2101 cggaagctca tatgtgcaga atggaaccgg aaacgcctac gaggaagagg ccaacaagca
     2161 gtcagtagga tccgcggcca cgaacttctc tctgttaaag caagcaggag acgtggaaga
     2221 aaaccccggt cctatgggca gatctgggga atcgctgttt aaagggccac gggattacaa
     2281 tcctatttca tccactatat gccatctgac caatgaatca gatggccata caacttccct
     2341 atacggaatc ggatttggac catttatcat cacgaataag catctgtttc gccggaataa
     2401 cggaactctc ctggtgcaga gcctccatgg tgtgtttaag gttaaaaaca caacaacatt
     2461 gcagcaacac ctcattgacg gccgggatat gatcataatc cgtatgccca aagatttccc
     2521 tccatttcct cagaagctca aatttcgtga gccacagagg gaagaacgga tatgtctggt
     2581 caccaccaat ttccagaccg ggggtagtgg gtccatcctc tggcacgaga tgtggcacga
     2641 aggtttagag gaggcaagcc gactgtactt cggagaacgc aacgtgaagg ggatgtttga
     2701 ggtgctggag cctcttcatg cgatgatgga acgggggcca caaacgctaa aggagacatc
     2761 ttttaaccag gcctacggtc gcgacctcat ggaggctcaa gagtggtgta gaaagtacat
     2821 gaagtcaggg aacgttaagg atctgctcca ggcctgggat ctttattacc atgtgttcag
     2881 aagaatatct aagcaggaag ggcgcggttc tttactgact tgtggggacg tagaggagaa
     2941 tcccgggccc ggggttcagg tcgaaacgat ttcacccggg gatggtcgca ccttcccgaa
     3001 acgagggcaa acttgcgtgg tccattacac cggcatgttg gaggatggca aaaaatttga
     3061 ttcttctcgg gaccggaaca agccctttaa attcatgctc gggaaacagg aagtcataag
     3121 aggctgggag gagggagttg cccaaatgag cgtcggtcag agagccaagc tcaccataag
     3181 cccagattat gcttacgggg ccacagggca tccaggtatc ataccccctc atgccacttt
     3241 ggtatttgac gtggaactgc tgaaattgga gggcggatca ggatctaaat ccatgagcag
     3301 catggtgagc gatacctctt gtacctttcc cagttctgat ggtatattct ggaaacactg
     3361 gattcagaca aaggacggac aatgcggatc tcctcttgtt tcgacgaggg atgggttcat
     3421 cgttggcatt cactccgcat ccaacttcac caacacaaac aactacttca cgtccgtgcc
     3481 taagaatttc atggaacttc ttacgaatca agaagcccag cagtgggttt ctggatggcg
     3541 cctcaacgcc gacagtgtgt tgtggggcgg acacaaagtg tttatggtga aaccagagga
     3601 accgttccag ccagtgaagg aggccacaca attaatgaac taaccgctcg agaattgttg
     3661 ttgttgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3721 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3781 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3841 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3901 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4021 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4141 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4261 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4321 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4381 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4441 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4561 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4621 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4681 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4741 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4801 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     4861 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     4921 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     4981 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     5041 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     5101 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     5161 tgaattgctg ccctctggtt atgtgtggga gggctaagcg ggactctggg gttcgaaatg
     5221 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5281 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5341 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5401 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5461 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5521 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5581 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5641 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6661 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6721 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6781 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6841 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6901 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6961 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7021 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7081 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7141 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7201 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7261 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7321 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7381 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7441 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7501 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7561 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7621 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7681 gtgccacctg acgtc
//
