LOCUS       V023285                 8112 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023285
VERSION     V023285
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2786
                     /label="Chgb(NM_007694)"
                     /note="Chgb(NM_007694)"
                     /gene="Chgb"
     CDS             2799..2864
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2888..3476
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3542..3590
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3792..4220
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4234..4563
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4630..5424
                     /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    5600..5721
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5770..5786)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5794..5810
                     /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(5818..5848)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5863..5884
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6172..6760)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6931..7791)
                     /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(7792..7896)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatccg ccaccatgca gccggctatg ctcctcggcc
      781 tcctgggcgc agcggctctg gccgctgtca gctcagctcc agtggataac agggatcaca
      841 atgaagaaat ggtgactcgc tgcattatcg aagtcctctc aaatgcccta tccaagtcca
      901 gtgttccaac gatcacccct gagtgccggc aagtcctgaa gaagagtgga aaagaggtca
      961 aaggtgaaga gaaaggtgaa aaccagaact cgaagtttga agtacggttg ctaagagacc
     1021 cagccgatgc ctcaggaacc cgctgggcct ccagtaggga ggatgcaggc gctccagtag
     1081 aagactctca aggccagacg aaggtaggca atgagaagtg gacagaagga ggaggacaca
     1141 gccgagaggg agtggatgat caagagagtc tccgtccctc caaccaacaa gcgtccaaag
     1201 aagcaaagat ataccattcg gaagagaggg tgggaaaaga gagggagaaa gaagaaggca
     1261 agatttaccc aatgggggag cacagggaag atgctggtga agagaagaaa cacattgaag
     1321 attctggaga gaagccgaac accttctcca acaaaagaag cgaggcttcc gctaagaaaa
     1381 aagatgagtc agtggccaga gcagatgcac actccatgga gctggaggag aagacacaca
     1441 gtagggagca aagcagccag gagagtggag aggagacaag gaggcaggag aaaccccagg
     1501 agcttaccga ccaggaccag agccaggaag aatcccagga aggcgaggaa ggtgaggaag
     1561 gcgaggaagg cgaggaaggc gaggaagact cagcctctga agtaaccaaa cgcaggccca
     1621 gacaccacca cgggagaagc gggtccaaca agtcctctta cgaagggcat cctctatctg
     1681 aggaaaggag accctctccc aaggagtcca aggaggcaga tgtggccaca gtccgtttag
     1741 gggaaaaaag gagtcaccat ctagctcact acagggcttc agaagaagaa cctgaatatg
     1801 gggaagaatc gagaagctat cggggactac agtaccgggg tagaggaagc gaagaggaca
     1861 gagccccaag gcctcggagt gaggagagtc aggaaaggga gtataagaga aaccaccccg
     1921 actctgaact ggaaagcaca gcaaacagac atggtgaaga aaccgaggaa gagaggagct
     1981 acgagggggc aaatggacgc caacacagag gcagggggcg ggagccagga gcccattctg
     2041 ctcttgacac cagagaagaa aaaaggctct tggatgaagg gcactaccct gttcgcgaaa
     2101 gcccaataga tacggcaaaa aggtacccac aaagcaaatg gcaagagcag gagaaaaact
     2161 acctcaacta tggcgaggaa ggggaccaag ggagatggtg gcagcaggaa gagcagctag
     2221 gaccggaaga aagcagggag gaagtaaggt ttccagacag acaatatgag ccctatccca
     2281 tcactgaaaa gaggaagagg ttaggggcgc tgttcaaccc gtactttgac cctctccagt
     2341 ggaagaatag tgactttgag aaaagaggca acccagatga cagttttctt gaggatgaag
     2401 gtgaagacag aaatggggtg accttgactg agaagaattc cttcccagag tacaactatg
     2461 actggtggga gagaaggccc ttctcagagg atgtgaattg gggatatgag aagagaagct
     2521 ttgccagggc ccctcagctc gacttgaaac ggcaatatga tggagtggcc gagttggacc
     2581 agctgcttca ctacaggaag aaggcagacg aatttcccga tttctacgac tcggaggagc
     2641 agatggggcc tcaccaggag gcaaacgatg aaaaggccag ggctgaccag agagttctga
     2701 cggcggaaga gaaaaaggaa ctggagaact tggctgcgat ggatctggaa ctccagaaga
     2761 tagcggaaaa gttcagccag cggggcctcg agtctagaga ctacaaagac catgacggtg
     2821 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2881 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2941 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3001 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3061 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3121 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3181 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3241 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3301 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3361 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3421 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3481 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3541 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3601 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3661 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3721 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3781 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3841 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3901 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3961 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4021 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4081 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4141 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4201 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4261 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4381 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4441 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4501 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4561 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4621 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4681 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4741 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4801 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4861 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4921 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4981 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5041 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5101 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5161 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5221 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5281 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5341 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5401 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5461 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5521 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5581 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5701 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5761 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5821 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5881 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5941 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6361 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6601 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7921 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7981 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8041 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8101 aatctgctta gg
//
