LOCUS       V033725                 8306 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033725
VERSION     V033725
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3745
                     /label="Itih2(NM_010582)"
                     /note="Itih2(NM_010582)"
                     /gene="Itih2"
     CDS             3752..3778
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3782..3808
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3812..3838
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3873..4097
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4143..4571
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4585..4914
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4981..5775
                     /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    5949..6070
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6119..6135)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6143..6159
                     /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(6167..6197)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6212..6233
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6521..7106)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7277..8137)
                     /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(8138..8242)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg cagcggcctg tgtgccttct catctggctg tttcttctgg aagcacaagc
      961 cttcgaaatc cccataaacg gaaactcaga atttgcagaa tacagtgatc ttgtggaact
     1021 ggccccagat aaactcccat ttgtgcaaga gaatggaaga catcagagaa gccttcctga
     1081 ggaatccgga gaggagacgg acactgttga tccagtgact ctttatagct acaaagtcca
     1141 gtccactatt acttctcggg tggccacaac taccatccag agcaaactgg tgaacaattc
     1201 cccattgcct cagagtgtcg tgttcgatgt tcagattccc aaaggagcct ttatctccaa
     1261 cttcaccatg actgtgaacg gcatgacatt tacaagctcc attaaagaga aaactgtggg
     1321 ccgagctctt tattcacagg caagagcaaa gggcaagacg gctggatggg taaggagcag
     1381 aactctggat atggagaact tcaacacaga ggtgaacatc ccgcctggag ccaaggtgca
     1441 gtttgaactt cattaccagg aggtgaagtg gaggaagctg ggatcctatg agcacaagat
     1501 tcatttgcag cccgggaagc tggccaaaca cttggaggtg aatgtgtgga ttatcgaacc
     1561 ccaaggaatg agatttcttc atgttcctga cacatttgaa ggccatttcc aaggcgtccc
     1621 tgtcatatct aaaggacaac agaaggcaca tgtatccttc aagcccacag tagcacaaca
     1681 aagaaaatgc cccaactgca cagagactgc agtgaacggg gagctggtgg tgatgtatga
     1741 tgtcaacaga gaagagaagg ccggggaact tgaggtgttt aatggatatt ttgttcactt
     1801 ctttgctcct gagaacctgg acccaatccc caagaacatc ctttttgtta ttgacgtcag
     1861 tggctccatg tgggggatta agatgaagca gactgtggag gcaatgaaga cgatattgga
     1921 tgacctaaga acagacgacc aattctctgt ggttgatttc aaccataatg ttcgaacctg
     1981 gagaaatgat ttagtttcag ctactaaaac acaaattgca gatgccaaga gatacattga
     2041 gaaaattcag cctagtggag gcacgaatat caacgaggca ctgcttcgag caattttcat
     2101 tttgaatgaa gccagtaaca tgggattgct gaaccccgac tcagtctctc tgatcatttt
     2161 ggtttctgat ggagatccaa ccgtagggga actaaaattg tccaaaattc agaaaaatgt
     2221 gaagcagagc atccaagata acatctccct gtttagtttg ggaatcggat ttgatgtcga
     2281 ctatgatttt ttgaagaggc tgtccaatga aaatcgtggt attgctcaac ggatctacgg
     2341 gaaccaggac acatcctctc agctcaagaa attttacaac caggtctcta ctccactgct
     2401 cagaaatgtt cagtttaact acccgcaggc atcagtaaca gatgtcaccc aaaacaattt
     2461 ccacaactac ttcggaggtt ctgagatcgt ggtggcaggg aaatttgatc ccagcaaact
     2521 gacagaagtc cagagcatca tcacggcaac ttcggctaac acagaattgg tcttggaaac
     2581 cctgagccag atggatgact tggaggagtt tctgtcaaag gacaagcatg cagaccctga
     2641 cttcaccaag aagctgtggg cctatctcac catcaaccaa ctactagcag agagaagtct
     2701 ggctcctact gctgccatca aaaggaaaat caccaaaaca atcctgcaga tgtctctaga
     2761 ccatcacatc gtgactccgc ttactgccat ggtgatagag aatgatgctg gagatgaacg
     2821 catgctggct gactccccgc cacaggacca ctcctgctgc tcaggcgcac tgtattatgg
     2881 caccaaggtt gcctcgggtc ccatcccatc atgggccaac ccatctccga caccaatgtc
     2941 ggccatgctt gcagtcggag cgaagccact tgagtctact ccacctactc atttgaatca
     3001 agtggaaaat gacccccact tcatcattta cctgcccaaa agcaaaagga atatttgttt
     3061 caatattgac tcagaacctg ggaaaatcct aagcctggtg tctgacccag aatcagggat
     3121 tgtagtcaac ggtcagctta tcggcgccaa gagggcagag aatggaaagc taagcaccta
     3181 ctttggaaaa ctggggtttt atttccaaaa ggaaggcatg aagatagaaa tcagcaccga
     3241 gaccatcacc ctgagctctg gctcctctac atctcgactg tcctggtccg acacagctca
     3301 tcttggaaac tcgagggtgc tcatctctgt gaagaaagaa aaatctgtga ctctcaccct
     3361 gaataaagag ctgttctttt ccgttctgct gcatcgtgtg tggaggaagc acccagttaa
     3421 cgtggacttc ctggggatct acgctccccc catagacaag ttctcaccta gagtgcatgg
     3481 actgttaggc cagttcatgc aggagccagc catccacatc ttcaatgaga gacccggaaa
     3541 ggagccggga aaacccgagg caagcatgga ggtgaaagga cataagctga ctgtcaccag
     3601 aggcctgcag aaggactaca ggacggatat cgtgtttgga acagacgttc cctgctggtt
     3661 tgtacacaac agtgggaaag gtttcatcga tggacattat aaggattatt ttgtgcctca
     3721 gctctacagc tttctcaagc ggcctctcga gtacccatac gacgtacctg attatgcaac
     3781 ttatccttat gatgtacctg attatgctag ctatccttat gatgtcccag actacgcatg
     3841 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3901 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3961 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4021 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4081 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4141 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4561 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4621 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4681 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4741 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4801 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4861 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4921 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4981 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5041 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5101 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5161 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5221 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5281 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5341 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5401 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5461 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5521 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5581 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5641 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5701 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5761 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5821 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5881 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5941 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6001 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6061 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6301 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7021 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7321 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7381 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7441 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7501 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7561 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7621 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7681 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7741 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7801 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7861 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7921 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7981 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8041 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8101 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8161 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8221 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8281 cgggagatct cccgatcccc tatggt
//
