LOCUS       V033251                 8228 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033251
VERSION     V033251
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             945..968
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             972..2933
                     /label="HSPA5(NM_005347)"
                     /note="HSPA5(NM_005347)"
                     /gene="HSPA5"
     polyA_signal    2987..3609
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3651..3948
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4002..4796
                     /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    5448..5582
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(5588..5606)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5620..5636)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5644..5660
                     /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(5668..5698)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5713..5734
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6022..6610)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6781..7641)
                     /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(7642..7746)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7773..8201
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcag atcgatctac catggactac aaagacgatg
      961 acgacaagct tatgaagctc tccctggtgg ccgcgatgct gctgctgctc agcgcggcgc
     1021 gggccgagga ggaggacaag aaggaggacg tgggcacggt ggtcggcatc gacctgggga
     1081 ccacctactc ctgcgtcggc gtgttcaaga acggccgcgt ggagatcatc gccaacgatc
     1141 agggcaaccg catcacgccg tcctatgtcg ccttcactcc tgaaggggaa cgtctgattg
     1201 gcgatgccgc caagaaccag ctcacctcca accccgagaa cacggtcttt gacgccaagc
     1261 ggctcatcgg ccgcacgtgg aatgacccgt ctgtgcagca ggacatcaag ttcttgccgt
     1321 tcaaggtggt tgaaaagaaa actaaaccat acattcaagt tgatattgga ggtgggcaaa
     1381 caaagacatt tgctcctgaa gaaatttctg ccatggttct cactaaaatg aaagaaaccg
     1441 ctgaggctta tttgggaaag aaggttaccc atgcagttgt tactgtacca gcctatttta
     1501 atgatgccca acgccaagca accaaagacg ctggaactat tgctggccta aatgttatga
     1561 ggatcatcaa cgagcctacg gcagctgcta ttgcttatgg cctggataag agggaggggg
     1621 agaagaacat cctggtgttt gacctgggtg gcggaacctt cgatgtgtct cttctcacca
     1681 ttgacaatgg tgtcttcgaa gttgtggcca ctaatggaga tactcatctg ggtggagaag
     1741 actttgacca gcgtgtcatg gaacacttca tcaaactgta caaaaagaag acgggcaaag
     1801 atgtcaggaa agacaataga gctgtgcaga aactccggcg cgaggtagaa aaggccaaac
     1861 gggccctgtc ttctcagcat caagcaagaa ttgaaattga gtccttctat gaaggagaag
     1921 acttttctga gaccctgact cgggccaaat ttgaagagct caacatggat ctgttccggt
     1981 ctactatgaa gcccgtccag aaagtgttgg aagattctga tttgaagaag tctgatattg
     2041 atgaaattgt tcttgttggt ggctcgactc gaattccaaa gattcagcaa ctggttaaag
     2101 agttcttcaa tggcaaggaa ccatcccgtg gcataaaccc agatgaagct gtagcgtatg
     2161 gtgctgctgt ccaggctggt gtgctctctg gtgatcaaga tacaggtgac ctggtactgc
     2221 ttgatgtatg tccccttaca cttggtattg aaactgtggg aggtgtcatg accaaactga
     2281 ttccaaggaa cacagtggtg cctaccaaga agtctcagat cttttctaca gcttctgata
     2341 atcaaccaac tgttacaatc aaggtctatg aaggtgaaag acccctgaca aaagacaatc
     2401 atcttctggg tacatttgat ctgactggaa ttcctcctgc tcctcgtggg gtcccacaga
     2461 ttgaagtcac ctttgagata gatgtgaatg gtattcttcg agtgacagct gaagacaagg
     2521 gtacagggaa caaaaataag atcacaatca ccaatgacca gaatcgcctg acacctgaag
     2581 aaatcgaaag gatggttaat gatgctgaga agtttgctga ggaagacaaa aagctcaagg
     2641 agcgcattga tactagaaat gagttggaaa gctatgccta ttctctaaag aatcagattg
     2701 gagataaaga aaagctggga ggtaaacttt cctctgaaga taaggagacc atggaaaaag
     2761 ctgtagaaga aaagattgaa tggctggaaa gccaccaaga tgctgacatt gaagacttca
     2821 aagctaagaa gaaggaactg gaagaaattg ttcaaccaat tatcagcaaa ctctatggaa
     2881 gtgcaggccc tcccccaact ggtgaagagg atacagcaga aaaagatgag ttgtaggaat
     2941 tcatcgatag atctgatatc ggtaccagtc gactctagag gatcccgggt ggcatccctg
     3001 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3061 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3121 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3181 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3241 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3301 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3361 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3421 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3481 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3541 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3601 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3841 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3901 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca
     3961 gcttgggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4801 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4861 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4921 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc
     4981 cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca
     5041 gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga
     5101 actgcaggag tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg
     5161 cctgatacag aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct
     5221 gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc
     5281 cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5341 cgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5401 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5461 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5521 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5581 tcaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     5641 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5701 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5761 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7741 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     7801 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7861 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7921 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7981 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     8041 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     8101 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     8161 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     8221 tacaattt
//
