LOCUS       62056_18320        6366 bp DNA     circular SYN 26-MAY-2020
DEFINITION  Cloning vector pP-anti-VEGF-scFV-MS2, complete sequence.
ACCESSION   MN996870
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  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 6366)
  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 6366)
  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..6366
                     /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             1685..1699
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     CDS             1730..1759
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1781..1807
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_RNA        1843..1861
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     polyA_signal    2161..2385
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2431..2859
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2873..3202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3269..3865
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4042..4175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4212..4228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4236..4252)
                     /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(4260..4290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4305..4326)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4614..5199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5373..6230)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6231..6335)
                     /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 cgcggccgca gatctgccac catgaatagc ttcagtacct ctgcttttgg
      901 cccggtagcc ttcagtctgg ggctactttt ggtactgccc gctgcttttc cagctccagt
      961 gcagttagtg gagagcggtg gggggttggt ccagccaggc ggaagcctac ggctgtcctg
     1021 tgctgcctca ggcttcacca taagcgacta ctggattcat tgggttcgac aagcccctgg
     1081 caaatgtctg gaatgggtcg ctggcatcac accggccggc gggtacactt actacgcaga
     1141 ttccgtgaaa ggacgcttta ccatcagtgc cgatacatct aaaaatacag cttatctcca
     1201 gatgaactca ctccgtgctg aggacacagc cgtttactat tgtgctcggt ttgtattttt
     1261 cctcccgtat gcaatggatt actggggaca aggcactctg gtgaccgttt cttctggcgg
     1321 agggggaagc ggaggaggtg ggtcgggtgg ggggggctca gacattcaga tgacccagtc
     1381 acctagcagc ttgtccgctt ccgtggggga tcgtgtgaca ataacatgtc gggcttcgca
     1441 ggacgtttct acagcagtgg cgtggtacca acagaaacct ggaaaggcac ctaagcttct
     1501 tatttacagc gcctcttttc tgtacagtgg agttcctagc cgatttagtg gctccgggtc
     1561 tgggacagac ttcaccttga ccatctcttc cctccagccc gaggatttcg ctacgtacta
     1621 ctgccagcaa cattatacga cccctccgac gtttggatgc ggaaccaaag ttgaaatcaa
     1681 gggggacgac gacgacaagt aatcccttga acctcctcgt tcgacctaag agcaaaagct
     1741 catttctgaa gaggacttga ccggttttct agctagctaa tacccatacg acgtaccaga
     1801 ttacgctgtg ggcatgagcg tgctcctcgg atcttaggga gcacatgagg atcacccatg
     1861 tgccacgagc gacatgagga tcacccatgt cgctcgtgtt cccgcgagca catgaggatc
     1921 acccatgtgc ctgtcgcgac atgaggatca cccatgtcgc gacagttcgc gccagcacat
     1981 gaggatcacc catgtgccgt gtgcgacatg aggatcaccc atgtcgcaca cgttggcccc
     2041 agcacatgag gatcacccat gtgccagacg cgacatgagg atcacccatg tcgcgtctgt
     2101 tgggggatcc gcgccgctcg agaattgttg ttgttgttta aacccgctga tcagcctcga
     2161 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2221 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2281 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2341 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2401 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2461 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2521 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2581 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2641 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2701 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2761 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2821 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2881 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2941 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3121 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3181 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3241 tcaagagaca ggatgaggat cgtttcgcat gaccgagtac aagcctaccg tgcgcctggc
     3301 cactcgcgat gatgtgcccc gcgccgtccg cactctggcc gccgctttcg ccgactaccc
     3361 cgctacccgg cacaccgtgg accccgaccg gcacatcgag cgtgtgacag agttgcagga
     3421 gctgttcctg acccgcgtcg ggctggacat cggcaaggtg tgggtagccg acgacggcgc
     3481 ggccgtggcc gtgtggacta cccccgagag cgttgaggcc ggcgccgtgt tcgccgagat
     3541 cggcccccga atggccgagc tgagcggcag ccgcctggcc gcccagcagc aaatggaggg
     3601 cctgcttgcc ccccatcgtc ccaaggagcc tgcctggttt ctggccactg taggagtgag
     3661 ccccgaccac cagggcaagg gcttgggcag cgccgtcgtg ttgcccggcg tagaggccgc
     3721 cgaacgcgcc ggtgtgcccg cctttctcga aacaagcgca ccaagaaacc ttccattcta
     3781 cgagcgcctg ggcttcaccg tgaccgccga tgtcgaggtg cccgagggac ctaggacctg
     3841 gtgtatgaca cgaaaacctg gcgcctaagc gggactctgg ggttcgaaat gaccgaccaa
     3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4021 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4141 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     5161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6361 gacgtc
//
