LOCUS       V032369                 6991 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032369
VERSION     V032369
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             706..1650
                     /label="Eif2s1(NM_019356)"
                     /note="Eif2s1(NM_019356)"
                     /gene="Eif2s1"
     CDS             1657..1683
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1687..1713
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1717..1743
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    1767..2355
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2421..2469
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2671..3099
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3113..3442
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3509..4303
                     /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    4479..4600
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4649..4665)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4673..4689
                     /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(4697..4727)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4742..4763
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5810..6670)
                     /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(6671..6775)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgcc gggtctaagt
      721 tgtagatttt atcaacacaa atttcctgag gtcgaagatg tagtgatggt gaatgtaaga
      781 tccattgctg aaatgggggc ctatgtcagc ttgttggaat ataataacat tgaaggcatg
      841 attcttctta gtgaattatc cagacgacgt atccgttcta taaacaaact gatccgaatt
      901 ggcagaaatg aatgtgtagt tgtcattaga gtggataaag aaaaaggata tatagatttg
      961 tcaaaaagaa gagtttctcc agaggaagca atcaaatgtg aagacaaatt cacaaaatcc
     1021 aaaactgttt atagcattct tcgccatgtt gctgaggtat tagagtatac caaggatgag
     1081 cagctggaaa gcctattcca gaggactgcc tgggtctttg atgacaagta caagagacct
     1141 ggatatggtg cctatgatgc ctttaagcat gcagtctcag acccatctat cttggatagt
     1201 ttagatttga atgaagatga aagagaagta ctcattaaca atatcaatag gcgtttgacc
     1261 ccacaagctg tcaagattcg agcagatatt gaggtagctt gctatggtta cgaaggcatt
     1321 gatgctgtaa aagaagccct gagagcaggt ttgaattgtt ctacagaaac catgcccatc
     1381 aagattaatc taatagctcc acccaggtat gtgatgacaa caacgaccct agagaggaca
     1441 gaaggactct ctgttctcaa tcaggctatg gcagtcatca aagaaaagat tgaggagaag
     1501 aggggagtgt tcaatgttca gatggagccc aaagtggtta cagatacaga tgagactgaa
     1561 cttgcaaggc agctggaacg gcttgagaga gaaaatgcag aagtggatgg agatgatgat
     1621 gcagaagaaa tggaagccaa agctgaagat ctcgagtacc catacgacgt acctgattat
     1681 gcaacttatc cttatgatgt acctgattat gctagctatc cttatgatgt cccagactac
     1741 gcatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     1801 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1861 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1921 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     1981 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2041 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2101 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2161 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2221 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2281 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2341 gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc
     2401 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     2461 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     2521 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     2581 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     2641 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2701 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2761 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2821 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2881 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2941 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3001 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     3061 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3361 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3421 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3481 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3661 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4021 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4321 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4381 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4441 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4621 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6781 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     6841 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6901 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     6961 gaccgacaat tgcatgaaga atctgcttag g
//
