LOCUS       V025732                 9353 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025732
VERSION     V025732
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             671..4012
                     /label="Eif2ak3(NM_010121)"
                     /note="Eif2ak3(NM_010121)"
                     /gene="Eif2ak3"
     CDS             4019..4045
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4049..4075
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4079..4105
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4129..4717
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4783..4831
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5033..5461
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5475..5804
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5871..6665
                     /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    6841..6962
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7011..7027)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7035..7051
                     /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(7059..7089)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7104..7125
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7413..8001)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8172..9032)
                     /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(9033..9137)
                     /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 taccgccacc atggagcgcg ccacccggcc cgggccgcgc gcgctgctgc tgcttctgtt
      721 cctgctgctg ggctgcgcgg cggggatctc ggcggtcgcg cccgcccgca gtttgcttgc
      781 tcccgcgtcg gagacagtgt ttggcttagg ggcagcggcc gccccgactt cggccgcgcg
      841 ggtgcctgcg gtggcaacgg ccgaagtgac cgtggaggac gccgaggcat tgccggctgc
      901 cgctggcgaa ccggagtcac gcgcgacgga gcccgatgac gacgtggaac tgcggcctcg
      961 cggcaggtcc ttggtaatca tcagcacttt agatggacga atcgctgcac tggatgccga
     1021 gaatgatggg aaaaagcagt gggatttgga cgtggggtct ggttccttgg tttcatctag
     1081 cctcagcaag ccagaggtgt ttgggaacaa gatgatcatc ccctccctgg atggagacct
     1141 cttccagtgg gaccgggacc gagagagcat ggaggccgtc cccttcacgg tggagtccct
     1201 gctcgaatct tcctacaagt ttggagatga tgttgttctg gttggaggga aatctctgac
     1261 tacatacgga ctcagtgctt acagtggaaa gctgaggtat atctgttctg ccttgggatg
     1321 tcgccgatgg gatagtgatg aaatggaaga agaggaagac atcttgcttc tgcagcgtac
     1381 gcagaagact gtgcgagctg tcgggcctcg aagcggcagt gagaagtgga atttcagtgt
     1441 tggccacttt gaacttcggt atattccaga catggaaact agagccggat tcattgaaag
     1501 cacctttaaa ccgggtggaa acaaagaaga ctctaaaatt atttcagatg tggaagaaca
     1561 agaagccacc atgctggaca cagtgataaa agtttccgtt gctgattgga aggtcatggc
     1621 gtttagtagg aagggaggcc gcctggaatg ggagtaccag ttttgtactc ccatcgcgtc
     1681 cgcctggctg gtgagggatg gcaaggtcat ccccatcagc ctgtttgatg atacaagtta
     1741 cacagccagc gaagaagcct tgggagacga agaagacatt gtagaggctg ctcggggagc
     1801 cacagagaac agcgtgtact tagggatgta cagaggccag ctgtacctgc agtcgtccgt
     1861 cagggtctca gaaaagttcc ctacaagccc aaaggccttg gagtctgtaa atggcgaaaa
     1921 tgcaattatt cctctgccga cgatcaaatg gaagccctta atccattctc cttctaggac
     1981 tcctgtcttg gttgggtctg atgaatttga caaatgtcta agtaatgata agtattccca
     2041 cgaagaatac agtaatggtg cactttcaat cctccagtat ccatacgata acggttacta
     2101 tctgccatac tacaagagag aaaggaataa gcggagcacg cagatcacag tcaggttcct
     2161 ggacagcccc cactacagca agaacatccg caagaaggac cctatcctcc tgctgcactg
     2221 gtggaaggag atattcggga cgatcctgct ttgcatcgta gccacgacct tcatcgtgcg
     2281 caggcttttc catcctcagc cccacaggca gcggaaggag tctgaaactc agtgccagac
     2341 tgaaagtaaa tacgactccg tgagtgccga tgtcagtgac aacagctgga atgacatgaa
     2401 gtactcagga tacgtatccc gatatctaac agattttgag ccaattcagt gcatgggtcg
     2461 tggtggcttt ggcgttgtct ttgaagctaa aaacaaagta gatgactgca attacgctat
     2521 caagaggatc cggctcccca acagggagtt ggcacgggag aaggtaatgc gggaagttaa
     2581 agccttggct aagctggaac acccaggcat tgtgaggtat ttcaacgcct ggctggaaac
     2641 cccaccagag aagtggcaag aagagatgga tgagatctgg ctcaaagacg aaagcacaga
     2701 ctggccgctc agctccccta gcccgatgga tgccccatct gttaagatcc gaaggatgga
     2761 tcctttctct acaaaagagc agatcgaagt catagctcct tctcctgaaa gaagtcggtc
     2821 tttctcggtg ggcatttcct gtggccagac aagctcatcg gagagccagt tctctcccct
     2881 ggagttctca gggacagact gcggagacaa cagtgactca gcggacgcag cctacaacct
     2941 ccaggacagt tgcctgacgg actgcgagga cgtggaagat ggcaccgtgg acggcaatga
     3001 cgagggacac tcctttgaac tttgtccgtc cgaagcttct ccctataccc ggtcaaggga
     3061 aggaacgtcc tcctccatag tgtttgagga ctctggctgc ggcaacgcgt ccagtaagga
     3121 ggagcccaga gggaaccggc tgcatgatgg caaccattat gttaataagc taactgatct
     3181 caagtgctcc agcagcaggt cttcttcaga agccaccacc ttgtctacct cccctaccag
     3241 gccaaccact ctaagcttgg atttcaccaa gaacactgtg ggccagctcc agcccagctc
     3301 ccccaaggtg tatctgtaca ttcagatgca gctgtgcagg aaggagaacc tcaaagactg
     3361 gatgaaccgg cgctgcagct tggaggaccg ggagcacggc gtgtgcctgc acatcttcct
     3421 gcagatcgca gaggcagtgg agttcctgca cagcaaggga ctcatgcaca gggacctcaa
     3481 gccttccaac atattcttca caatggatga tgtggtcaag gttggggact ttggactggt
     3541 gactgctatg gaccaagatg aagaagagca gactgtactg actccaatgc cagcctatgc
     3601 tacgcacacg ggacaagtag ggaccaagct atacatgagc ccagagcaga ttcatggaaa
     3661 caactactcc cataaagtgg acatcttctc tttaggcttg attctgtttg aactcctcta
     3721 cccattcagc acccagatgg aacgagtccg gattttaact gatgtcagaa atctcaagtt
     3781 tcctctactg ttcactcaga aatatcccca agagcatatg atggttcaag acatgctctc
     3841 tccatccccc acggagcggc ctgaagccac agacatcatt gaaaatgcca tatttgagaa
     3901 cttggagttt cccgggaaaa cggttctgag acagcggtcc cgctccatga gttcatctgg
     3961 aacaaaacat tccagacagc ccagctgctc gtacagccca ctgcctggca acctcgagta
     4021 cccatacgac gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta
     4081 tccttatgat gtcccagact acgcatagtg agggcccgat atctcgacaa tcaacctctg
     4141 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4201 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4261 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4321 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4381 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4441 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4501 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     4561 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4621 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4681 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     4741 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4801 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4861 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4921 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4981 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     5041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5161 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     5221 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5281 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5341 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5401 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5461 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5521 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5641 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5701 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5761 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5821 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5881 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5941 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     6001 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     6061 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6121 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6181 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6241 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6301 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6361 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     6421 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6481 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6541 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6601 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6661 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     6721 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6781 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6841 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6901 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6961 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     7621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9181 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     9241 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     9301 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
