LOCUS       V015809                 7572 bp    DNA     circular SYN 26-MAY-2020
DEFINITION  Exported.
ACCESSION   V015809
VERSION     V015809
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7572)
  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 7572)
  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 7572)
  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..7572
                     /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="TVMV site"
                     /note="tobacco vein mottling virus (TVMV) NIa 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             2246..2602
                     /gene="rpl7ae"
                     /label="Large ribosomal subunit protein eL8"
                     /note="Large ribosomal subunit protein eL8 from
                     Archaeoglobus fulgidus (strain ATCC 49558 / DSM 4304 / JCM
                     9628 / NBRC 100126 / VC-16). Accession#: O29494"
     CDS             2672..2695
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     polyA_signal    3568..3792
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3838..4266
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4280..4609
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4676..5071
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    5248..5381
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5418..5434)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5442..5458)
                     /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(5466..5496)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5511..5532)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5820..6405)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6579..7436)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7437..7541)
                     /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 gagacggtta gattccagag tggctcagga 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 gatctatgta tgtgagattc gaagttcctg aagatatgca
     2281 gaacgaggcg ctaagcctcc tggaaaaggt gcgtgaatca gggaaagtga aaaaaggaac
     2341 gaatgaaact actaaagcag tggaacgagg tcttgccaaa ctggtgtaca ttgccgagga
     2401 tgtcgaccca cccgaaatcg tcgcccattt gcctcttctg tgtgaagaga agaacgtgcc
     2461 ctatatctac gtcaaatcta aaaacgatct gggacgcgcg gtgggcatcg aggtgccatg
     2521 cgcgtccgcg gccatcatta atgaggggga gctgcggaag gagctgggga gtcttgtgga
     2581 gaagattaag ggcctccaga agggatccgc ggacgaaatg gaggaatgca gtcaacactt
     2641 gcccggcgca ggatcctccg gagacattat ggattataaa gacgatgacg acaaaggatc
     2701 ttctggcact ggctccggta gcggtacaag tgcgcccata acagcttacg ctcagcagac
     2761 aaggggactt ttgggatgca taattacatc acttaccgga agggacaaaa atcaggtaga
     2821 aggagaggtc cagattgtga gcactgctac tcaaaccttt ctggccacat gtattaatgg
     2881 agtgtgttgg gccgtttatc acggggcagg cacccggaca atcgcaagtc ccaagggccc
     2941 cgtgattcaa atgtacacca acgtggacca ggatctagta ggatggcccg ccccgcaggg
     3001 ctctcgcagc ctgacccctt gtacctgcgg aagttcagac ctgtaccttg tcacaagaca
     3061 tgcggacgtt atcccggtcc gccgaagggg tgactccaga ggaagcttgc tgagccccag
     3121 gcctatctcg tatttgaagg gctctagtgg tggaccgctc ctctgcccag ccgggcatgc
     3181 agtcggtcta ttccgtgccg ctgtgtgcac aaggggcgtc gcaaaagctg tggatttcat
     3241 ccccgtcgag aatttggaga ccactatgcg ctcccccgtt tttacggaca attcttcgcc
     3301 ccccgcagtg acgctgaccc accctattac aaagatcgat accaaataca tcatgacgtg
     3361 catgtcagcg gatctggagt ggacaagtac ctgggtgttg gtaggcgggg tgctcgctgc
     3421 acttgctgct tactgcctga gcacagggtg cgtcgtgatc gttgggcgca ttgtgctatc
     3481 agggaaaccc gcgatgccag accgagaagt cctgtactaa ccgctcgaga attgttgttg
     3541 ttgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3601 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3661 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3721 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3781 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4201 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4381 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4441 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4501 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4561 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4621 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatggc
     4681 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     4741 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     4801 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     4861 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     4921 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     4981 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     5041 attgctgccc tctggttatg tgtgggaggg ctaagcggga ctctggggtt cgaaatgacc
     5101 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5161 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5221 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5341 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5401 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5461 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5521 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5581 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5641 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5701 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5761 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5821 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5881 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5941 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6001 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6061 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6121 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6181 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6241 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6301 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     6361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7561 ccacctgacg tc
//
