LOCUS       V045223                 7891 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045223
VERSION     V045223
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3381
                     /label="Vcp(NM_009503)"
                     /note="Vcp(NM_009503)"
                     /gene="Vcp"
     CDS             3388..3453
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3491..3715
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3761..4189
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4203..4532
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4599..5393
                     /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    5567..5688
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5737..5753)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5761..5777
                     /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(5785..5815)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5830..5851
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6139..6724)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6895..7755)
                     /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(7756..7860)
                     /gene="bla"
                     /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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcct ctggagccga ttcaaaaggt gatgatttat caacagccat tctcaaacag
     1021 aagaaccgac ccaatcggtt aattgttgat gaagccatca atgaagataa cagcgtggtg
     1081 tccttgtccc agcccaagat ggatgaactg cagttgttcc gaggtgacac ggtgttgcta
     1141 aaaggaaaga aaagacggga agctgtatgc attgttcttt ctgatgacac gtgttctgat
     1201 gagaagattc gaatgaatag agttgttcgg aataacctcc gagttcgcct aggagatgtc
     1261 atcagcatcc agccatgccc tgatgtaaag tatggcaaac gtatccacgt tctacccatc
     1321 gatgacacag tggaaggcat cactggcaat ctctttgagg tataccttaa gccgtacttc
     1381 ctggaagctt atcggcccat ccgtaaagga gatatttttc ttgtccgggg tgggatgcgt
     1441 gctgtggagt tcaaagttgt agagacagat cccagccctt actgtattgt tgctccagac
     1501 acagtgatcc actgtgaggg ggagccaatc aagcgagagg atgaggagga atccttgaat
     1561 gaagtaggct atgatgacat cggtggttgc aggaagcagc tagctcagat aaaggagatg
     1621 gtggagctgc cactgagaca tcctgcgctc tttaaggcga ttggtgtaaa gcctcctcgg
     1681 ggaatcttgt tgtatgggcc tcctgggaca gggaagaccc tgattgctcg agctgtggca
     1741 aatgaaactg gagccttctt ctttctgatc aatggtcctg aaatcatgag caaattggct
     1801 ggtgagtctg agagcaacct tcgtaaagcc tttgaggaag ctgaaaagaa tgctcctgct
     1861 atcatcttca tcgatgagct tgatgccatt gcacccaaaa gagagaaaac tcatggggaa
     1921 gtggagcgtc gcatcgtgtc tcagttgttg accctcatgg atggcctaaa gcagagagca
     1981 catgtgatag ttatggcagc aaccaataga cccaacagca ttgacccagc cctacggcga
     2041 tttggtcgct ttgacagaga ggtagatatt ggaatacctg atgctacagg acgtttggag
     2101 attcttcaga tccataccaa gaacatgaaa ctggcagatg atgtggactt ggaacaggta
     2161 gccaatgaga ctcatggtca tgttggtgct gatttggcag ccctatgttc agaggctgct
     2221 ctgcaggcca tccggaaaaa aatggacctc attgacctag aagatgagac cattgatgct
     2281 gaggtcatga attccctggc agttactatg gatgacttcc ggtgggcttt gagtcaaagc
     2341 aacccatcag cacttcggga aactgtggta gaggtgccac aagtaacctg ggaagatatt
     2401 ggaggcctgg aggatgtcaa acgtgagctt caggagttgg ttcagtatcc tgtggaacat
     2461 ccagacaaat tcctcaaatt tggcatgact ccctccaaag gcgttctttt ctatggacct
     2521 cctggctgtg ggaaaacctt actggctaaa gccattgcta atgaatgcca ggccaacttc
     2581 atctccatca agggtcctga gctgcttacc atgtggtttg gggaatctga ggccaatgtc
     2641 cgggaaattt ttgacaaggc acggcaagct gccccctgtg tactcttctt tgatgagtta
     2701 gattcaattg ccaaggctcg aggtggtaat attggagatg gtggtggagc tgctgaccga
     2761 gtcatcaatc agatcctgac agaaatggat ggcatgtcta caaaaaagaa tgtgtttatc
     2821 attggagcta ccaacaggcc tgacatcatt gatcctgcta tcctaagacc tggccgtcta
     2881 gatcagctca tttatatccc acttcctgat gagaagtccc gtgttgccat cctaaaagcc
     2941 aatctgcgaa agtccccagt tgccaaggat gtggatttgg agttcctggc taagatgact
     3001 aatggctttt ctggagctga tttgacagaa atttgccaac gggcttgtaa actggccatt
     3061 cgtgaatcta ttgagagtga gattaggcga gaacgagaga ggcagacaaa tccatcggct
     3121 atggaggtag aagaggacga tccagtgcct gagatccgca gagatcactt tgaggaagcc
     3181 atgcgttttg cccgacgttc tgtcagcgat aatgacattc ggaagtatga aatgttcgcc
     3241 cagacactgc agcagagtcg aggttttggc agcttcagat tcccttcagg gaaccagggt
     3301 ggagctggtc ccagccaggg cagtggaggt ggcacaggtg gcagtgtgta cacagaagac
     3361 aatgacgatg acctgtatgg cctcgaggac tacaaagacc atgacggtga ttataaagat
     3421 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta aacccgctga
     3481 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3541 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3601 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3661 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3721 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3781 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3841 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3901 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3961 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4021 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4081 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4141 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4201 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4381 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4441 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4501 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4561 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5101 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5401 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5581 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5701 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5761 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5821 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5881 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6661 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7861 cacatttccc cgaaaagtgc cacctgacgt c
//
