LOCUS       V015921                 6720 bp    DNA     circular SYN 26-MAY-2020
DEFINITION  Exported.
ACCESSION   V015921
VERSION     V015921
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6720)
  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 6720)
  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 6720)
  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..6720
                     /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             1277..2131
                     /gene="CNOT7"
                     /label="CCR4-NOT transcription complex subunit 7"
                     /note="CCR4-NOT transcription complex subunit 7 from Homo
                     sapiens. Accession#: Q9UIV1"
     CDS             2150..2170
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             2183..2656
                     /label="ecDHFR"
                     /note="E. coli dihydrofolate reductase"
     polyA_signal    2716..2940
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2986..3414
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3428..3757
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3824..4219
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    4396..4529
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4566..4582)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4590..4606)
                     /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(4614..4644)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4659..4680)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4968..5553)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5727..6584)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6585..6689)
                     /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 atcgctatgc cggctgccac cgttgaccac tcacaaagga tctgcgaagt
     1321 gtgggcctgt aaccttgatg aggagatgaa gaagatcaga caggtcatta ggaagtacaa
     1381 ctatgttgct atggacacag agttcccagg agtggtcgca aggcctattg gcgagttccg
     1441 gagcaatgca gattatcagt atcagttgtt gaggtgtaac gtggatctct taaaaatcat
     1501 tcagctcgga cttacattca tgaacgaaca gggcgaatat ccccccggca cttctacatg
     1561 gcagttcaat ttcaagttta atctgacaga ggacatgtat gcccaggatt cgatcgagct
     1621 gctcacaact agcggaatcc agtttaagaa gcacgaggaa gaaggcatcg aaacccagta
     1681 ctttgcagag cttttgatga cgtctggggt ggtgctctgt gagggcgtca agtggctcag
     1741 ctttcatagt ggttatgatt ttggttatct catcaagatc ctgactaaca gtaatctccc
     1801 agaagaggag ctagactttt tcgagatact ccggctcttc tttcccgtta tatatgacgt
     1861 gaaatatctc atgaaaagct gtaagaacct caaaggaggg cttcaggaag tggcggagca
     1921 gctggagctt gaaagaattg gaccacagca tcaggccggg tctgactcac tgctcaccgg
     1981 gatggccttc ttcaaaatgc gcgagatgtt tttcgaagat cacattgacg atgccaagta
     2041 ctgcgggcat ctgtacgggc ttggcagcgg aagctcatat gtgcagaatg gaaccggaaa
     2101 cgcctacgag gaagaggcca acaagcagtc agtaggatct ggtgggtccg agaatcttta
     2161 tttccaatcc ggatctggca gtatcagtct gatcgccgcc cttgctgtgg attacgtgat
     2221 tgggatggag aatgccatgc catggaacct tcctgctgat cttgcttggt ttaaacgtaa
     2281 cacactgaac aagccggtga ttatgggtcg tcatacctgg gaatctattg gccgtcctct
     2341 gccggggcgg aagaatatta tcctcagtag tcagccgtct accgacgata gggtgacttg
     2401 ggttaagtca gttgatgagg ccattgctgc ttgtggcgac gtgcctgaaa taatggttat
     2461 tggcggaggg cgggtcatcg aacagtttct tcccaaggca cagaaactgt accttactca
     2521 catagacgcg gaggtggagg gggacaccca tttccctgat tatgagccag atgactggga
     2581 gtcagtattt agtgagttcc acgatgccga tgcccagaac agtcactcgt attgttttga
     2641 aattttggag cgccgaggat ccgcgtaacc gctcgagaat tgttgttgtt gtttaaaccc
     2701 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2761 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2821 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2881 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2941 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3001 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3061 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3121 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3181 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3241 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3301 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3361 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3421 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3481 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3601 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3661 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3721 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3781 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatggcca agcctttgtc
     3841 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     3901 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     3961 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     4021 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     4081 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     4141 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     4201 tggttatgtg tgggagggct aagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4261 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4321 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4381 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4441 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4501 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4561 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4621 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4681 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4741 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     5521 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5581 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5641 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5701 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5761 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5821 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5881 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5941 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6001 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6061 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6121 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6181 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6241 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6301 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6361 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6421 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6481 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6541 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6601 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6661 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
