LOCUS       V027565                 7509 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027565
VERSION     V027565
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2189
                     /label="Ica1(NM_010492)"
                     /note="Ica1(NM_010492)"
                     /gene="Ica1"
     CDS             2196..2261
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2285..2873
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2939..2987
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3189..3617
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3631..3960
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4027..4821
                     /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    4997..5118
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5167..5183)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5191..5207
                     /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(5215..5245)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5260..5281
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5569..6157)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6328..7188)
                     /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(7189..7293)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtc aggacacaaa tgttattcct
      781 gggagttgca ggaccggttt gctcaagata agtcagttgt caataagatg cagcagaaat
      841 attgggagac caagcaggcc tttatcaaag ccacggggaa gaaggaagat gaacatgtgg
      901 ttgcttctga tgcagacctg gatgccaagc tggagctgtt tcactcaatc cagagaacct
      961 gcctggactt gtctaaagca atcgtgctct atcaaaagag aatatgtttc ttgtctcaag
     1021 aggaaaatga actgggaaag tttcttcgat ctcaaggctt ccaagacaaa acccgagcag
     1081 gaaaaatgat gcaagccacg ggcaaggccc tctgcttttc ctcccagcaa aggttggcct
     1141 tgagaaaccc tttgtgtcga tttcaccaag aagtagaaac ttttagacat cgggccatct
     1201 ctgatacctg gctgacagtg aaccgcatgg agcagtacag gacagaatac agaggagcgt
     1261 tgttgtggat gaaggatgtg tctcaggaac ttgatccaga cctctacaag caaatggaga
     1321 agttcaggaa ggtgcagaca caagtccgcc ttgcgaaaaa aaactttgac aagttgaaga
     1381 tggatgtgtg tcaaaaggtg gatcttcttg gagcaagcag atgtaacctc ttatctcaca
     1441 tgctagcaac ataccagacc actctgctcc acttttggga gaaaacttct cacaccatgg
     1501 cagccattca tgagagcttc aaaggttatc aaccatatga attcacaacg ttaaagagct
     1561 tacaagaccc catgaagaag ctagttgaga aagaagggaa gaagacctcc tggagggaaa
     1621 accgggaggc tgtggcacca gagccgaggc agttaatttc tttggaggat gagcacaaag
     1681 attcatctgc ttataagact gaagagggaa caagcgtttt gtcttccgta gacaaaggtt
     1741 ctgtacatga cacatgctca ggacccatag atgaactatt agacgggaaa cccgaggaag
     1801 cgtgcctggg tcccacggca gggaccccag aacctgaaag tggggacaag gatgacctct
     1861 tgctgttgaa tgagatcttc agcacttcct gcctggatga gggagagttc agcagagagt
     1921 gggctgcagt gtttggagat gaccagctaa aggagccagc acccatgggg gcccaaggag
     1981 agccagaccc caagccccag ataggctccg gattccttcc gtcacagctt ttagaccaaa
     2041 atatgaaaga tctccaggcc tctctgcaag agcctgccaa ggctgcctcg gacctgactg
     2101 cctggttcag cctctttgct gacctcgacc ccttatcaaa ccctgatgct gttgggaaaa
     2161 ctgataaaga acacgaattg ctcaatgcag gatccgacta caaagaccat gacggtgatt
     2221 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2281 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2341 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2401 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2461 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2521 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2581 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2641 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2701 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2761 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2821 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2881 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2941 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3001 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3061 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3121 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3181 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3421 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3481 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3541 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3601 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3961 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4801 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4981 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5041 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5101 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5161 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5221 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5281 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5341 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5401 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5461 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5521 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5581 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5641 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5701 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5761 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5821 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5881 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5941 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6001 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6061 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7321 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7381 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7441 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7501 ctgcttagg
//
