LOCUS       V018355                 7475 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018355
VERSION     V018355
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2932
                     /label="CHFR(NM_001161344)"
                     /note="CHFR(NM_001161344)"
                     /gene="CHFR"
     CDS             2972..3037
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3075..3299
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3345..3773
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3787..4116
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4183..4977
                     /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    5151..5272
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5321..5337)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5345..5361
                     /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(5369..5399)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5414..5435
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5723..6308)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6479..7339)
                     /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(7340..7444)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggagcggc ccgaggaagg
      961 caagcagtcg ccgccgccgc agccctgggg acggctcctg cgtctgggcg cggaggaggg
     1021 cgagccgcac gtcctcctga ggaagcggga gtggaccatc gggcggagac gaggttgcga
     1081 cctttccttc cccagcaata aactggtctc tggagatcac tgtagaattg tagtggatga
     1141 aaaatcaggt caggtgacac tggaagatac cagcaccagt ggaacagtga ttaacaagct
     1201 gaaggttgtt aagaagcaga catgcccttt acagactggg gatgtcatct acttggtgta
     1261 caggaagaat gaaccggaac acaacgtggc atacctctat gaatctttaa gtgaaaagca
     1321 aggcatgaca caagaatcct ttgaagctaa caaggaaaat gtgttccatg ggaccaaaga
     1381 tacctcaggt gcaggtgcag ggcgaggggc cgatccccgg gtccctccgt cgtcgcccgc
     1441 cactcaggtg tgctttgagg aaccacagcc atcaacatcg acgtcagacc tcttccccac
     1501 agcctcggcc tcttccacgg agccttctcc tgcagggcga gagcgttcct ccagttgtgg
     1561 gtctgggggt ggtggcatct cccctaaagg aagtggtccc tctgtggcaa gtgatgaagt
     1621 ctccagcttt gcctcagctc tcccagacag aaagactgcg tccttttcgt cgttggaacc
     1681 ccaggatcag gaggatttgg agcccgtgaa gaagaaaatg agaggagatg gggaccttga
     1741 cctgaacggg cagttgttgg tcgcacaacc gcgtagaaat gcccaaaccg tccacgagga
     1801 cgtcagagca gcggctggga agccagacaa gatggaggag acgctgacat gcatcatctg
     1861 ccaggacctg ctgcacgact gcgtgagttt gcagccctgc atgcacacgt tctgcgcggc
     1921 ttgctactcg ggctggatgg agcgctcgtc cctgtgtcct acctgccgct gtcccgtgga
     1981 gcggatctgt aaaaaccaca tcctcaacaa cctcgtggaa gcatacctca tccagcatcc
     2041 agacaagagt cgcagtgaag aagatgtgca aagtatggat gccaggaata aaatcactca
     2101 agacatgctg cagcccaaag tcaggcggtc tttttctgat gaagaaggga gttcagagga
     2161 cctgctggag ctgtcagacg ttgacagtga gtcctcagac attagccagc catacgtcgt
     2221 gtgccggcag tgtcctgagt acagaaggca ggcggcgcag cctccccact gcccagcacc
     2281 cgagggcgag ccaggagccc cacaggccct gggggatgca ccctccacgt ccgtcagcct
     2341 gacgacagca gtccaggatt acgtgtgccc tctgcaagga agccacgccc tgtgcacctg
     2401 ctgcttccag cccatgcccg accggagagc ggagcgcgag caggacccgc gtgtcgcccc
     2461 tcagcagtgt gcggtctgcc tgcagccttt ctgccacctg tactggggct gcacccggac
     2521 cggctgctac ggctgcctgg ccccgttttg tgagctcaac ctgggtgaca agtgtctgga
     2581 cggcgtgctg aacaacaaca gctacgagtc agacatcctg aagaattacc tggcaaccag
     2641 aggtttgaca tggaaaaaca tgttgaccga gagcctcgtg gctctccagc ggggagtgtt
     2701 tctgctgtct gattacagag tcacgggaga caccgttctg tgttactgct gtggcctgcg
     2761 cagcttccgt gagctgacct atcagtatcg gcagaacatt cctgcttccg agttgccagt
     2821 ggccgtaaca tcccgtcctg actgctactg gggccgtaac tgccgcactc aggtgaaagc
     2881 tcaccacgcc atgaaattca atcatatctg tgaacagaca aggttcaaaa acgaattctg
     2941 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa
     3001 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg
     3061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5041 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5101 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5161 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5221 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5281 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5341 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5401 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5461 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5521 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5581 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5641 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6241 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7441 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
