LOCUS       V018623                 8924 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018623
VERSION     V018623
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      619..628
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1846..4725
                     /label="MET(NM_001324402)"
                     /note="MET(NM_001324402)"
                     /gene="MET"
     CDS             4732..4752
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQG"
                     /label="TEV site"
     CDS             4765..4848
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     CDS             4870..5589
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="mEGFP"
     polyA_signal    5712..5833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5840..6295)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6322..6426
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        6428..6785
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6820..7614
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7846..7893
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      8222..8810
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatggag tgtaaagagg cactagcaaa gtccgagatg
      661 aatgtgaata tgaagtatca gcttcccaac ttcaccgcgg aaacacccat ccagaatgtc
      721 attctacatg agcatcacat tttccttggt gccactaact acatttatgt tttaaatgag
      781 gaagaccttc agaaggttgc tgagtacaag actgggcctg tgctggaaca cccagattgt
      841 ttcccatgtc aggactgcag cagcaaagcc aatttatcag gaggtgtttg gaaagataac
      901 atcaacatgg ctctagttgt cgacacctac tatgatgatc aactcattag ctgtggcagc
      961 gtcaacagag ggacctgcca gcgacatgtc tttccccaca atcatactgc tgacatacag
     1021 tcggaggttc actgcatatt ctccccacag atagaagagc ccagccagtg tcctgactgt
     1081 gtggtgagcg ccctgggagc caaagtcctt tcatctgtaa aggaccggtt catcaacttc
     1141 tttgtaggca ataccataaa ttcttcttat ttcccagatc atccattgca ttcgatatca
     1201 gtgagaaggc taaaggaaac gaaagatggt tttatgtttt tgacggacca gtcctacatt
     1261 gatgttttac ctgagttcag agattcttac cccattaagt atgtccatgc ctttgaaagc
     1321 aacaatttta tttacttctt gacggtccaa agggaaactc tagatgctca gacttttcac
     1381 acaagaataa tcaggttctg ttccataaac tctggattgc attcctacat ggaaatgcct
     1441 ctggagtgta ttctcacaga aaagagaaaa aagagatcca caaagaagga agtgtttaat
     1501 atacttcagg ctgcgtatgt cagcaagcct ggggcccagc ttgctagaca aataggagcc
     1561 agcctgaatg atgacattct tttcggggtg ttcgcacaaa gcaagccaga ttctgccgaa
     1621 ccaatggatc gatctgccat gtgtgcattc cctatcaaat atgtcaacga cttcttcaac
     1681 aagatcgtca acaaaaacaa tgtgagatgt ctccagcatt tttacggacc caatcatgag
     1741 cactgcttta ataggacact tctgagaaat tcatcaggct gtgaagcgcg ccgtgatgaa
     1801 tatcgaacag agtttaccac agctttgcag cgcgttgact tattcatggg tcaattcagc
     1861 gaagtcctct taacatctat atccaccttc attaaaggag acctcaccat agctaatctt
     1921 gggacatcag agggtcgctt catgcaggtt gtggtttctc gatcaggacc atcaacccct
     1981 catgtgaatt ttctcctgga ctcccatcca gtgtctccag aagtgattgt ggagcataca
     2041 ttaaaccaaa atggctacac actggttatc actgggaaga agatcacgaa gatcccattg
     2101 aatggcttgg gctgcagaca tttccagtcc tgcagtcaat gcctctctgc cccacccttt
     2161 gttcagtgtg gctggtgcca cgacaaatgt gtgcgatcgg aggaatgcct gagcgggaca
     2221 tggactcaac agatctgtct gcctgcaatc tacaaggttt tcccaaatag tgcacccctt
     2281 gaaggaggga caaggctgac catatgtggc tgggactttg gatttcggag gaataataaa
     2341 tttgatttaa agaaaactag agttctcctt ggaaatgaga gctgcacctt gactttaagt
     2401 gagagcacga tgaatacatt gaaatgcaca gttggtcctg ccatgaataa gcatttcaat
     2461 atgtccataa ttatttcaaa tggccacggg acaacacaat acagtacatt ctcctatgtg
     2521 gatcctgtaa taacaagtat ttcgccgaaa tacggtccta tggctggtgg cactttactt
     2581 actttaactg gaaattacct aaacagtggg aattctagac acatttcaat tggtggaaaa
     2641 acatgtactt taaaaagtgt gtcaaacagt attcttgaat gttatacccc agcccaaacc
     2701 atttcaactg agtttgctgt taaattgaaa attgacttag ccaaccgaga gacaagcatc
     2761 ttcagttacc gtgaagatcc cattgtctat gaaattcatc caaccaaatc ttttattagt
     2821 ggtgggagca caataacagg tgttgggaaa aacctgaatt cagttagtgt cccgagaatg
     2881 gtcataaatg tgcatgaagc aggaaggaac tttacagtgg catgtcaaca tcgctctaat
     2941 tcagagataa tctgttgtac cactccttcc ctgcaacagc tgaatctgca actccccctg
     3001 aaaaccaaag cctttttcat gttagatggg atcctttcca aatactttga tctcatttat
     3061 gtacataatc ctgtgtttaa gccttttgaa aagccagtga tgatctcaat gggcaatgaa
     3121 aatgtactgg aaattaaggg aaatgatatt gaccctgaag cagttaaagg tgaagtgtta
     3181 aaagttggaa ataagagctg tgagaatata cacttacatt ctgaagccgt tttatgcacg
     3241 gtccccaatg acctgctgaa attgaacagc gagctaaata tagagtggaa gcaagcaatt
     3301 tcttcaaccg tccttggaaa agtaatagtt caaccagatc agaatttcac aggattgatt
     3361 gctggtgttg tctcaatatc aacagcactg ttattactac ttgggttttt cctgtggctg
     3421 aaaaagagaa agcaaattaa agatctgggc agtgaattag ttcgctacga tgcaagagta
     3481 cacactcctc atttggatag gcttgtaagt gcccgaagtg taagcccaac tacagaaatg
     3541 gtttcaaatg aatctgtaga ctaccgagct acttttccag aagatcagtt tcctaattca
     3601 tctcagaacg gttcatgccg acaagtgcag tatcctctga cagacatgtc ccccatccta
     3661 actagtgggg actctgatat atccagtcca ttactgcaaa atactgtcca cattgacctc
     3721 agtgctctaa atccagagct ggtccaggca gtgcagcatg tagtgattgg gcccagtagc
     3781 ctgattgtgc atttcaatga agtcatagga agagggcatt ttggttgtgt atatcatggg
     3841 actttgttgg acaatgatgg caagaaaatt cactgtgctg tgaaatcctt gaacagaatc
     3901 actgacatag gagaagtttc ccaatttctg accgagggaa tcatcatgaa agattttagt
     3961 catcccaatg tcctctcgct cctgggaatc tgcctgcgaa gtgaagggtc tccgctggtg
     4021 gtcctaccat acatgaaaca tggagatctt cgaaatttca ttcgaaatga gactcataat
     4081 ccaactgtaa aagatcttat tggctttggt cttcaagtag ccaaaggcat gaaatatctt
     4141 gcaagcaaaa agtttgtcca cagagacttg gctgcaagaa actgtatgct ggatgaaaaa
     4201 ttcacagtca aggttgctga ttttggtctt gccagagaca tgtatgataa agaatactat
     4261 agtgtacaca acaaaacagg tgcaaagctg ccagtgaagt ggatggcttt ggaaagtctg
     4321 caaactcaaa agtttaccac caagtcagat gtgtggtcct ttggcgtgct cctctgggag
     4381 ctgatgacaa gaggagcccc accttatcct gacgtaaaca cctttgatat aactgtttac
     4441 ttgttgcaag ggagaagact cctacaaccc gaatactgcc cagacccctt atatgaagta
     4501 atgctaaaat gctggcaccc taaagccgaa atgcgcccat ccttttctga actggtgtcc
     4561 cggatatcag cgatcttctc tactttcatt ggggagcact atgtccatgt gaacgctact
     4621 tatgtgaacg taaaatgtgt cgctccgtat ccttctctgt tgtcatcaga agataacgct
     4681 gatgatgagg tggacacacg accagcctcc ttctgggaga catcaggtac cgagaatctc
     4741 tacttccagg gacatatggc tagctggagc catccgcagt ttgaaaaagg tggtggtagc
     4801 ggtggtggtt caggtggtag tgcatggtca caccctcagt ttgagaaact ggatccaccg
     4861 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4921 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4981 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5041 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     5101 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5161 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5221 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5281 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5341 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5401 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5461 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     5521 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5581 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     5641 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5701 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5761 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5821 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5881 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5941 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6001 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6061 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6121 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6181 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6241 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     6301 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6361 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6421 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     6481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6601 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6721 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6781 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6841 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6901 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6961 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     7021 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     7081 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     7141 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     7201 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     7261 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     7321 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     7381 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7441 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7501 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7561 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7621 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     7681 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7741 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     7801 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     7861 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7921 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7981 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     8041 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     8101 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8161 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8221 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8281 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8341 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     8401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8521 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8701 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8761 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8821 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8881 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
