LOCUS       V033670                 7046 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033670
VERSION     V033670
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             941..2536
                     /label="ICAM1(NM_000201)"
                     /note="ICAM1(NM_000201)"
                     /gene="ICAM1"
     CDS             2543..2608
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2646..2870
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2916..3344
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3358..3687
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3754..4548
                     /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    4722..4843
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4892..4908)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4916..4932
                     /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(4940..4970)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4985..5006
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5294..5879)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6050..6910)
                     /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(6911..7015)
                     /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 atccgccacc atggctccca gcagcccccg
      961 gcccgcgctg cccgcactcc tggtcctgct cggggctctg ttcccaggac ctggcaatgc
     1021 ccagacatct gtgtccccct caaaagtcat cctgccccgg ggaggctccg tgctggtgac
     1081 atgcagcacc tcctgtgacc agcccaagtt gttgggcata gagaccccgt tgcctaaaaa
     1141 ggagttgctc ctgcctggga acaaccggaa ggtgtatgaa ctgagcaatg tgcaagaaga
     1201 tagccaacca atgtgctatt caaactgccc tgatgggcag tcaacagcta aaaccttcct
     1261 caccgtgtac tggactccag aacgggtgga actggcaccc ctcccctctt ggcagccagt
     1321 gggcaagaac cttaccctac gctgccaggt ggagggtggg gcaccccggg ccaacctcac
     1381 cgtggtgctg ctccgtgggg agaaggagct gaaacgggag ccagctgtgg gggagcccgc
     1441 tgaggtcacg accacggtgc tggtgaggag agatcaccat ggagccaatt tctcgtgccg
     1501 cactgaactg gacctgcggc cccaagggct ggagctgttt gagaacacct cggcccccta
     1561 ccagctccag acctttgtcc tgccagcgac tcccccacaa cttgtcagcc cccgggtcct
     1621 agaggtggac acgcagggga ccgtggtctg ttccctggac gggctgttcc cagtctcgga
     1681 ggcccaggtc cacctggcac tgggggacca gaggttgaac cccacagtca cctatggcaa
     1741 cgactccttc tcggccaagg cctcagtcag tgtgaccgca gaggacgagg gcacccagcg
     1801 gctgacgtgt gcagtaatac tggggaacca gagccaggag acactgcaga cagtgaccat
     1861 ctacagcttt ccggcgccca acgtgattct gacgaagcca gaggtctcag aagggaccga
     1921 ggtgacagtg aagtgtgagg cccaccctag agccaaggtg acgctgaatg gggttccagc
     1981 ccagccactg ggcccgaggg cccagctcct gctgaaggcc accccagagg acaacgggcg
     2041 cagcttctcc tgctctgcaa ccctggaggt ggccggccag cttatacaca agaaccagac
     2101 ccgggagctt cgtgtcctgt atggcccccg actggacgag agggattgtc cgggaaactg
     2161 gacgtggcca gaaaattccc agcagactcc aatgtgccag gcttggggga acccattgcc
     2221 cgagctcaag tgtctaaagg atggcacttt cccactgccc atcggggaat cagtgactgt
     2281 cactcgagat cttgagggca cctacctctg tcgggccagg agcactcaag gggaggtcac
     2341 ccgcaaggtg accgtgaatg tgctctcccc ccggtatgag attgtcatca tcactgtggt
     2401 agcagccgca gtcataatgg gcactgcagg cctcagcacg tacctctata accgccagcg
     2461 gaagatcaag aaatacagac tacaacaggc ccaaaaaggg acccccatga aaccgaacac
     2521 acaagccacg cctcccctcg aggactacaa agaccatgac ggtgattata aagatcatga
     2581 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2881 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3241 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3601 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3661 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3721 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4561 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4621 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4681 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4861 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5821 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7021 ttccccgaaa agtgccacct gacgtc
//
