LOCUS       V019318                 9634 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019318
VERSION     V019318
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             907..5124
                     /label="FAM135B(NM_015912)"
                     /note="FAM135B(NM_015912)"
                     /gene="FAM135B"
     CDS             5131..5196
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5234..5458
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5504..5932
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5946..6275
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6342..7136
                     /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    7310..7431
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7480..7496)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7504..7520
                     /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(7528..7558)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7573..7594
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7882..8467)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8638..9498)
                     /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(9499..9603)
                     /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 gccaccatgt ctgaaataca aggaacggtt gagttttcgg tagagctaca taaattttat
      961 aatgtggatc tctttcagag agggtattac cagatccgag tgaccttgaa ggtgtcttca
     1021 aggatccccc acagactgag tgcctccatc gctgggcaga cagagagcag cagcctgcat
     1081 tcagcctgtg tccatgacag caccgtgcac agccgggtct ttcagatctt ataccggaat
     1141 gaagaggtac ccataaatga tgctgtggtc ttccgagttc atttactctt gggtggtgaa
     1201 aggatggaag acgcactgag tgaagtagat tttcaactca aggtggatct gcactttacg
     1261 gacagtgaac agcagttgag ggatgtggct ggggcaccga tggtcagcag ccgaacgctt
     1321 ggcctgcact tccacccccg gaatggtctg caccaccagg tcccggtcat gttcgactat
     1381 ttccacctgt ctgtgatctc ggtgaccgtc catgctgccc tggtggctct gcagcagcca
     1441 ttgatcagtt ttactcgtcc aggaagaggc tcctggcttg gtaaaggtgg cccagacacc
     1501 ggacaagaac agtctatcat ttctctggaa aacttggtct ttggagctgg gtactgcaag
     1561 ccgacttcct cagagggaag cttctacatc acctctgaga actgcatgca gcacgcacac
     1621 aagtggcacc gagacctgtg cctgttgctc ctccacgctt accggggtct ccgtctccac
     1681 ttcctggtga tcatgcggga catcccagag ctgccacaca cggagctgga ggcccttgct
     1741 gttgaagaaa cactttctca gctgtgctca gagctacaga tgttgaacaa tccagagaag
     1801 atcgctgagc agataagcaa ggatctggcc tggctcacgt cccacatgat gactctgtgg
     1861 acccagttcc tggacacagt cactctgcac tcccaagtga ccacttatct cacccaggaa
     1921 caccacaccc tgagggtccg aaggttttct gaggccttct tttacatgga gcaccaaaaa
     1981 cttgcagtcc tgacatttca ggagaatctg atacagacgc acagccagct gtccctggat
     2041 atccggaact cggagtacct cactagcatg cccccgctgc ctgcagagtg cctggacatc
     2101 gacggcgatt ggaacaccct gccggtgatc tttgaggaca gatacgtgga ctgccctgcg
     2161 acagggcata acttgagtgt ttatcctaat tttgatgttc cagtgacaag tcctacaata
     2221 atgaatctga aagacaagga agataactgt atggtaaata gcaatttatc ttttagggaa
     2281 gaccttgtct tgtctaccat aaaaccatcc caaatggatt ctgatgaaga agttataagg
     2341 tgtccagagc caggtgagaa tgtggccaca caaaatcata tggacatgtg ctctgaatct
     2401 caggtgtata tatcaattgg tgaatttcaa aacaaagcag gtgtgcctga agatgaatgt
     2461 tggactggcc aaacatctga tgctgggaca tatccagtgg cagatgtgga tacttctaga
     2521 aggagtccag gtccagagga tggacaggcc ccagtgctga cctacattga cgtaaaatct
     2581 agcaataaga acccctccag agctgaaccc ctggtggcct tcaatgctca gcatgagagt
     2641 aggagctcta gagataagta tggattagac aggactgggc taagcaaagt ggtagtaggt
     2701 ggaagccacc aaaatgccat ctcttcagac aaaacaactc tccatgaatt aagtactcta
     2761 ggaaagggaa tagatcaaga ggggaagatg gtgctgctaa gcttgaaact caccccctct
     2821 gagccctgtg atccactaag ttctaccctg agggagccct tagatattag gtcttcccta
     2881 aaggactctg acacagaaga gcaggaggaa ctctcagtgc tatccggggt catcaagaga
     2941 tcttcatcca tcatatctga ttcaggcatt gagagtgagc caagctccgt cgcctggtca
     3001 gaggcccgaa gcagggctct ggagttgccc agtgatcggg aagtcttgca cccgtttgtt
     3061 cgaagacatg ccctccaccg gaactcccta gagggtggac acacagaaag taacacaagt
     3121 ttgccaagcg gcatccaggc ttctctcacc tccattagct ctttaccttt tgaggaggat
     3181 gagcgggagg tggcactcac taagttaacc aagtctgtat ctgctcccca catcagtagc
     3241 ccagaggagg ctgctgaaga tgcggacacc aagcagcaag atggaggttt tgctgaacct
     3301 tcagatatgc acagcaagag ccaaggttcc ccaggatctt gctctcaact ttgtggtgac
     3361 tctggaacag atgctggagc agaccatccc ctggtggaga tagttttaga tgctgacaac
     3421 cagcagggcc ccggatacat agacatcccc aaagggaaag ggaagcagtt tgatgctcaa
     3481 ggacactgtc ttcctgatgg caggactgag aacactccag gtgttgaaac caaaggtctt
     3541 aatttaaaaa taccacgcgt catagcactt gaaaacccca ggaccagatc tcttcataga
     3601 gcacttgagg aaaccccaaa gggcatgcct aaagacttga atgtgggtca gcaagctctt
     3661 tccaacagtg gcatctcaga ggttgagggt ctctctcaac atcaggtgcc tgaattgagc
     3721 tgtacgtcag ctgctgatgc catcaacagg aactcaacag gccagcaaag ccaaagcggt
     3781 tcaccttgca ttatggatga cacagcattt aatagaggag tgaatgcctt cccggaggct
     3841 aaacataaag caggcactgt gtgccccact gtgacccatt ccgttcattc ccaggttttg
     3901 aaaaaccaag agctgaaggc aggcacttcc atcatggggt cccatctgac ttctgcagag
     3961 acctttactc tggacagcct gaaggctgtg gaggttgtga acttatctgt gtcttgcact
     4021 gccacctgtc tccctttctc atctgtgccc aaggagaccc ctgccagggc tggattctct
     4081 tccaaacaga ccctgtttcc catcacccat cagcctttgg gatcctttgg agttgtttct
     4141 acccattcca gcacgttgga tgaggaagtc agtgagagga tgtttagttt ttatcaggcc
     4201 aaagaaaaat ttaaaaaaga actgaagatt gaaggatttc tgtacagtga cttaactgta
     4261 ctagcttctg atataccata tttcccacca gaggaagagg aagaaaattt ggaagatgga
     4321 attcacctgg ttgtctgtgt ccatggcctg gatgggaaca gtgcagacct ccggctggta
     4381 aagactttca tagaactggg gctccctgga ggaaaactgg acttcctaat gtctgaaaag
     4441 aatcagatgg acacatttgc agattttgat actatgacgg atcggttatt ggatgaaatc
     4501 attcaacaca ttcagttgta caacctctcc atatcccgaa ttagcttcat tggccattct
     4561 cttggcaaca tcatcatccg atcggtcctc acacggcccc ggttccggta ttacctcaac
     4621 aaactccaca cgttcctgtc actctctggg cctcacctgg gaaccctgta caacaacagc
     4681 accctggtta gtacaggctt atggctcatg cagaaactga agaaatccgg gtctctactg
     4741 cagctgacct tcagggataa tgctgatttg cgcaaatgtt tcctctacca actaagccaa
     4801 aaaacagggc tgcagtattt taaaaacgtc gtgctggttg cttctcccca agaccgttat
     4861 gtgccatttc attcagccag gattgaaatg tgtaaaactg ccctcaaaga cagacacaca
     4921 gggccagttt atgcagaaat gatcaacaac ctcctgggcc ctctggttga agccaaggac
     4981 tgcactttaa tccgacacaa cgtgttccac gccctgccca acactgccaa caccctgatc
     5041 ggccgagccg ctcacatcgc tgtgctggat tcagaactct tcctggagaa gtttttcttg
     5101 gtggcaggac tcaactactt caagctcgag gactacaaag accatgacgg tgattataaa
     5161 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     5221 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5281 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5341 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5401 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     5461 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5521 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5581 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5641 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5701 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5761 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5821 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5881 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5941 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6001 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6061 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     6121 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6181 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     6241 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     6301 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6361 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6421 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6481 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     6541 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6601 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6661 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6721 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6781 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6841 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6901 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6961 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     7021 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     7081 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     7141 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7201 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7261 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     7321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7441 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     8341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8401 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9601 gcgcacattt ccccgaaaag tgccacctga cgtc
//
