LOCUS       V016359                 8022 bp    DNA     circular SYN 26-MAY-2020
DEFINITION  Exported.
ACCESSION   V016359
VERSION     V016359
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8022)
  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 8022)
  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 8022)
  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..8022
                     /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..2683
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             2696..3052
                     /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             3122..3145
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     polyA_signal    4018..4242
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4288..4716
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4730..5059
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5126..5521
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    5698..5831
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5868..5884)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5892..5908)
                     /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(5916..5946)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5961..5982)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6270..6855)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7029..7886)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7887..7991)
                     /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 tccgcggcca cgaacttctc
     2641 tctgttaaag caagcaggag acgtggaaga aaaccccggt cctatgggca gatctatgta
     2701 tgtgagattc gaagttcctg aagatatgca gaacgaggcg ctaagcctcc tggaaaaggt
     2761 gcgtgaatca gggaaagtga aaaaaggaac gaatgaaact actaaagcag tggaacgagg
     2821 tcttgccaaa ctggtgtaca ttgccgagga tgtcgaccca cccgaaatcg tcgcccattt
     2881 gcctcttctg tgtgaagaga agaacgtgcc ctatatctac gtcaaatcta aaaacgatct
     2941 gggacgcgcg gtgggcatcg aggtgccatg cgcgtccgcg gccatcatta atgaggggga
     3001 gctgcggaag gagctgggga gtcttgtgga gaagattaag ggcctccaga agggatccgc
     3061 ggacgaaatg gaggaatgca gtcaacactt gcccggcgca ggatcctccg gagacattat
     3121 ggattataaa gacgatgacg acaaaggatc ttctggcact ggctccggta gcggtacaag
     3181 tgcgcccata acagcttacg ctcagcagac aaggggactt ttgggatgca taattacatc
     3241 acttaccgga agggacaaaa atcaggtaga aggagaggtc cagattgtga gcactgctac
     3301 tcaaaccttt ctggccacat gtattaatgg agtgtgttgg gccgtttatc acggggcagg
     3361 cacccggaca atcgcaagtc ccaagggccc cgtgattcaa atgtacacca acgtggacca
     3421 ggatctagta ggatggcccg ccccgcaggg ctctcgcagc ctgacccctt gtacctgcgg
     3481 aagttcagac ctgtaccttg tcacaagaca tgcggacgtt atcccggtcc gccgaagggg
     3541 tgactccaga ggaagcttgc tgagccccag gcctatctcg tatttgaagg gctctagtgg
     3601 tggaccgctc ctctgcccag ccgggcatgc agtcggtcta ttccgtgccg ctgtgtgcac
     3661 aaggggcgtc gcaaaagctg tggatttcat ccccgtcgag aatttggaga ccactatgcg
     3721 ctcccccgtt tttacggaca attcttcgcc ccccgcagtg acgctgaccc accctattac
     3781 aaagatcgat accaaataca tcatgacgtg catgtcagcg gatctggagt ggacaagtac
     3841 ctgggtgttg gtaggcgggg tgctcgctgc acttgctgct tactgcctga gcacagggtg
     3901 cgtcgtgatc gttgggcgca ttgtgctatc agggaaaccc gcgatgccag accgagaagt
     3961 cctgtactaa ccgctcgaga attgttgttg ttgtttaaac ccgctgatca gcctcgactg
     4021 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4081 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4141 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4201 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4261 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4681 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4741 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4801 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4921 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4981 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5041 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5101 agagacagga tgaggatcgt ttcgcatggc caagcctttg tctcaagaag aatccaccct
     5161 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     5221 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     5281 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     5341 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     5401 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     5461 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     5521 ctaagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5581 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5641 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6781 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     6841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
