LOCUS       V021153                 7511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021153
VERSION     V021153
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2158
                     /label="ATG14(NM_014924)"
                     /note="ATG14(NM_014924)"
                     /gene="ATG14"
     CDS             2198..2263
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2287..2875
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2941..2989
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3191..3619
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3633..3962
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4029..4823
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4999..5120
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5169..5185)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5193..5209
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5217..5247)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5262..5283
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5571..6159)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6330..7190)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7191..7295)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcgtc tcccagtggg aagggagccc gggcgctgga
      721 ggctcctggc tgcgggcccc ggccgctcgc ccgggacctg gtggactccg tggacgatgc
      781 ggaggggctg tacgtggctg tggagcgctg cccgctgtgc aacactaccc gccggcggct
      841 gacctgcgcc aaatgcgttc agagcggcga tttcgtctac ttcgacggcc gcgaccggga
      901 gaggtttatc gacaagaagg aaaggttaag ccgacttaag agcaagcaag aagaatttca
      961 gaaagaagtg ttaaaagcta tggaaggaaa atggataaca gatcagttga gatggaaaat
     1021 aatgtcctgc aagatgagga ttgaacagtt aaaacaaaca atatgtaaag gaaatgaaga
     1081 aatggagaaa aattctgaag gccttctcaa aaccaaggaa aagaatcaga agctttacag
     1141 tcgagcacaa cggcaccaag agaaaaagga gaagattcag aggcataatc gcaaacttgg
     1201 tgacctggta gaaaaaaaga ccattgactt aagaagtcat tatgagcgtc tggcaaatct
     1261 tcgacgatcc catatattag agctcacctc tgtcattttt ccaatcgagg aagtaaagac
     1321 gggtgtgaga gaccccgcag atgtgtcttc agagagtgac agtgccatga cctccagcac
     1381 tgtgagcaag cttgctgaag cccggaggac aacttacctc tcaggacgat gggtctgtga
     1441 cgatcacaac ggagacacca gcattagcat tacagggcct tggattagcc tccctaacaa
     1501 tggggactac tctgcctact acagctgggt ggaggagaag aaaacaaccc aggggcctga
     1561 catggagcag agtaaccctg cctacaccat cagtgctgcg ctgtgctatg caactcagct
     1621 ggtcaacatt ctgtctcata tacttgatgt aaatcttccc aaaaagctct gcaacagtga
     1681 attttgtggc gaaaatctaa gcaagcagaa atttactcga gcagtgaaga aactgaatgc
     1741 aaatattctt tacctttgtt tttctcagca tgtaaattta gatcaattac aaccactgca
     1801 taccctcagg aatctaatgt acctggtcag tccaagctct gaacacctag gcaggtcagg
     1861 gccctttgaa gtacgagcag accttgagga gtccatggaa tttgtggatc ccggagttgc
     1921 tggagaatca gatgagagcg gagatgagcg cgtcagcgat gaagaaaccg acctgggcac
     1981 agactgggag aacttgccta gtccccggtt ttgtgatatc ccttcccagt ctgtggaagt
     2041 ctcccagagt cagagcaccc aggcgtcccc acccatcgcg agcagcagtg caggtgggat
     2101 gatctcctct gcagcagcct cggtgacctc ctggtttaaa gcttacactg gacaccgtga
     2161 attctgcaga tatccagcac agtggcggcc gctcgaggac tacaaagacc atgacggtga
     2221 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     2281 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2341 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2401 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2461 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2521 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2581 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2641 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     2701 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2761 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2821 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2881 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2941 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     3001 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     3061 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     3121 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     3181 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3241 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3301 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     3361 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3421 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3481 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3541 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3601 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3661 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3901 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3961 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4861 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4921 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4981 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5101 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5161 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5221 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5761 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6001 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7321 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     7381 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7441 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7501 atctgcttag g
//
