LOCUS       V022178                 6891 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022178
VERSION     V022178
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             647..1561
                     /label="Ubb(NM_011664)"
                     /note="Ubb(NM_011664)"
                     /gene="Ubb"
     CDS             1562..1579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    1667..2255
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2321..2369
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2571..2999
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3013..3342
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3409..4203
                     /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    4379..4500
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4549..4565)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4573..4589
                     /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(4597..4627)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4642..4663
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4951..5539)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5710..6570)
                     /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(6571..6675)
                     /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 gccaccatgc agatcttcgt
      661 gaagaccctg accggcaaga ccatcaccct agaggtggag cccagtgaca ccatcgagaa
      721 cgtgaaggcc aagatccagg ataaagaggg catcccccct gaccagcaga ggctgatctt
      781 tgccggcaag cagctggaag atggccgcac cctctctgat tacaacatcc agaaggagtc
      841 aaccctgcac ctggtccttc gcctgagagg tggcatgcag atcttcgtga agaccctgac
      901 cggcaagacc atcaccctgg aggtggagcc cagtgacacc atcgagaacg tgaaggccaa
      961 gatccaggat aaagagggca tcccccctga ccagcagagg ctgatctttg ccggcaagca
     1021 gctggaagat ggccgcaccc tctctgatta caacatccag aaagagtcaa ccctgcacct
     1081 ggtcctccgt ctgaggggtg gcatgcagat cttcgtgaag accctgactg gcaagaccat
     1141 caccctggag gtggagccca gtgacaccat cgagaacgtg aaggccaaga tccaggataa
     1201 agagggcatc ccccctgacc agcagaggct gatctttgcc ggcaagcagc tggaagatgg
     1261 ccgcaccctc tctgattaca acatccagaa ggagtcaacc ctgcacctgg tccttcgcct
     1321 gagaggtggc atgcagatct tcgtgaagac cctgaccggc aagaccatca ccttggaggt
     1381 ggagcccagt gacaccatcg agaatgtgaa ggccaagatc caggataaag agggcatccc
     1441 ccctgaccag cagaggctga tctttgccgg caagcagcta gaagatggcc gcactctctc
     1501 tgattacaac atccagaaag agtcgaccct gcacctggtc ctccgtctga ggggtggcta
     1561 tcaccatcac catcaccatt aggatccact agtccagtgt ggtggaattc tgcagatatc
     1621 cagcacagtg gcggccgctc gagtctagag ggcccgatat ctcgacaatc aacctctgga
     1681 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1741 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1801 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     1861 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     1921 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     1981 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2041 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2101 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2161 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2221 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2281 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2341 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2401 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2461 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2521 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     2581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2701 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     2761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2941 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3001 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3061 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3361 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4201 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4261 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4321 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4381 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4441 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4501 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6721 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     6781 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6841 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
