LOCUS       62056_11990        6465 bp DNA     circular SYN 26-MAY-2020
DEFINITION  Cloning vector pH-K-turns_TVMVp, complete sequence.
ACCESSION   MN996848
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  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 6465)
  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 6465)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Oncolytics 17, 70-82 (2020)"
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6465
                     /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             1724..1735
                     /codon_start=1
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
                     /translation="WELQ"
     polyA_signal    1822..2046
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2092..2520
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2534..2863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2930..3964
                     /codon_start=1
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     polyA_signal    4141..4274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4311..4327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4335..4351)
                     /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(4359..4389)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4404..4425)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4713..5298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5472..6329)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6330..6434)
                     /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 ggcgtgatcc gaaaggtgac ccggatctgg ggcgtgatcc
      901 gaaaggtgac ccggatccac cggtcgccac catgggcaga tctagcaagg cacttctcaa
      961 aggtgtccgc gacttcaatc ccatctctgc ttgtgtctgc ctgcttgaga atagtagcga
     1021 tgggcatagc gagaggctgt ttggaatcgg cttcggccca tacattattg caaatcagca
     1081 ccttttccgg agaaataacg gggagctgac cattaaaaca atgcacggtg aatttaaggt
     1141 taagaatagt acccagctcc agatgaaacc cgtggaaggg cgggatataa tcgtgattaa
     1201 aatggcgaaa gattttccgc catttccgca gaagctgaaa ttccgtcagc ctactatcaa
     1261 agacagagtc tgcatggtgt caactaactt ccaacagggc ggttcaggat ctaaatcggt
     1321 cagctcctta gtgtcagaga gttctcatat agtgcacaaa gaggacacat ccttttggca
     1381 acactggatc actactaagg atggacagtg tggaagtccg ttagtgtcaa tcatcgatgg
     1441 gaatattttg ggtatacatt ccctcactca tacaacgaat gggtcaaact acttcgttga
     1501 gttccccgag aaattcgttg ccacttacct ggacgccgct gacggatggt gtaaaaactg
     1561 gaaattcaat gccgacaaaa tatcttgggg tagctttacg ttggtagagg acgctcccga
     1621 ggacgatttt atggcgaaaa aaacagttgc cgcaataatg gacagcggtg gcgcccttaa
     1681 gaaggaactg caagcaaata aaaaggagct ggctcagctg aagtgggaac tgcaagcact
     1741 aaagaaagaa ctggcccagt ccggatccgc gtaaccgctc gagaattgtt gttgttgttt
     1801 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2041 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2881 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgaaaaagcc
     2941 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     3001 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     3061 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3121 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     3181 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     3241 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     3301 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     3361 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     3421 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     3481 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     3541 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     3601 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     3661 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     3721 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     3781 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     3841 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     3901 actcgccgat agtggaaacc gacgccccag cactcgtccg gatcgggaga tgggggaggc
     3961 taactgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4021 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4081 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4141 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4201 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4261 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4321 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4381 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4441 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5221 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     5281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
