LOCUS       62056_11975        6591 bp DNA     circular SYN 26-MAY-2020
DEFINITION  Cloning vector pH-AFP-riboswitch_TVMVp, complete sequence.
ACCESSION   MN996865
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6591)
  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 6591)
  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 6591)
  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..6591
                     /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             1850..1861
                     /codon_start=1
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
                     /translation="WELQ"
     polyA_signal    1948..2172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2218..2646
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2660..2989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3056..4090
                     /codon_start=1
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     polyA_signal    4267..4400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4437..4453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4461..4477)
                     /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(4485..4515)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4530..4551)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4839..5424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5598..6455)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6456..6560)
                     /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 ggagagcgga agcgtgctgg gctccgaaca gcggaaggtg
      901 gttcgaagct ggggctttgg acataaccca gaggtcgatg gatccggatc tggggagagc
      961 ggaagcgtgc tgggctccga acagcggaag gtggttcgaa gctggggctt tggacataac
     1021 ccagaggtcg atggatccgg atccaccggt cgccaccatg ggcagatcta gcaaggcact
     1081 tctcaaaggt gtccgcgact tcaatcccat ctctgcttgt gtctgcctgc ttgagaatag
     1141 tagcgatggg catagcgaga ggctgtttgg aatcggcttc ggcccataca ttattgcaaa
     1201 tcagcacctt ttccggagaa ataacgggga gctgaccatt aaaacaatgc acggtgaatt
     1261 taaggttaag aatagtaccc agctccagat gaaacccgtg gaagggcggg atataatcgt
     1321 gattaaaatg gcgaaagatt ttccgccatt tccgcagaag ctgaaattcc gtcagcctac
     1381 tatcaaagac agagtctgca tggtgtcaac taacttccaa cagggcggtt caggatctaa
     1441 atcggtcagc tccttagtgt cagagagttc tcatatagtg cacaaagagg acacatcctt
     1501 ttggcaacac tggatcacta ctaaggatgg acagtgtgga agtccgttag tgtcaatcat
     1561 cgatgggaat attttgggta tacattccct cactcataca acgaatgggt caaactactt
     1621 cgttgagttc cccgagaaat tcgttgccac ttacctggac gccgctgacg gatggtgtaa
     1681 aaactggaaa ttcaatgccg acaaaatatc ttggggtagc tttacgttgg tagaggacgc
     1741 tcccgaggac gattttatgg cgaaaaaaac agttgccgca ataatggaca gcggtggcgc
     1801 ccttaagaag gaactgcaag caaataaaaa ggagctggct cagctgaagt gggaactgca
     1861 agcactaaag aaagaactgg cccagtccgg atccgcgtaa ccgctcgaga attgttgttg
     1921 ttgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1981 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2041 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2101 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2161 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2401 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2521 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2641 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2821 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2941 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3001 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgaa
     3061 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3121 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3181 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3241 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3301 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3361 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3421 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3481 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3541 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3601 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3661 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     3721 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     3781 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     3841 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     3901 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     3961 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4021 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccggatc gggagatggg
     4081 ggaggctaac tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4261 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4381 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4441 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4501 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4561 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     5401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5701 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
