LOCUS       V021144                 7968 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021144
VERSION     V021144
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"
     regulatory      895..904
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             946..3501
                     /label="MLXIPL(NM_032951)"
                     /note="MLXIPL(NM_032951)"
                     /gene="MLXIPL"
     promoter        complement(3520..3538)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3564..3788
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3834..4262
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4276..4606
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4673..5467
                     /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    5641..5762
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5811..5827)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5835..5851
                     /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(5859..5889)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5904..5925
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6213..6801)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6972..7832)
                     /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(7833..7937)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttgccacc
      901 atggactaca aggacgacga tgataagggt accgagctcg gatccatggc cggcgcgctg
      961 gcaggtctgg ccgcgggctt gcaggtcccg cgggtcgcgc ccagcccaga ctcggactcg
     1021 gacacagact cggaggaccc gagtctccgg cgcagcgcgg gcggcttgct ccgctcgcag
     1081 gtcatccaca gcggtcactt catggtgtcg tcgccgcaca gcgactcgct gccccggcgg
     1141 cgcgaccagg aggggtccgt ggggccctcc gacttcgggc cgcgcagtat cgaccccaca
     1201 ctcacacgcc tcttcgagtg cttgagcctg gcctacagtg gcaagctggt gtctcccaag
     1261 tggaagaatt tcaaaggcct caagctgctc tgcagagaca agatccgcct gaacaacgcc
     1321 atctggaggg cctggtatat ccagtatgtg aagcggagga agagccccgt gtgtggcttc
     1381 gtgacccccc tgcaggggcc tgaggctgat gcgcaccgga agccggaggc cgtggtcctg
     1441 gaggggaact actggaagcg gcgcatcgag gtggtgatgc gggaatacca caagtggcgc
     1501 atctactaca agaagcggct ccgtaagccc agcagggaag atgacctcct ggcccctaag
     1561 caggcggaag gcaggtggcc gccgccggag caatggtgca aacagctctt ctccagtgtg
     1621 gtccccgtgc tgctggggga cccagaggag gagccgggtg ggcggcagct cctggacctc
     1681 aattgctttt tgtccgacat ctcagacact ctcttcacca tgactcagtc cggcccttcg
     1741 cccctgcagc tgccgcctga ggatgcctac gtcggcaatg ctgacatgat ccagccggac
     1801 ctgacgccac tgcagccaag cctggatgac ttcatggaca tctcagattt ctttaccaac
     1861 tcccgcctcc cacagccgcc catgccttca aacttcccag agccccccag cttcagcccc
     1921 gtggttgact ccctcttcag cagtgggacc ctgggcccag aggtgccccc ggcttcctcg
     1981 gccatgaccc acctctctgg acacagccgt ctgcaggctc ggaacagctg ccctggcccc
     2041 ttggactcca gcgccttcct gagttctgat ttcctccttc ctgaagaccc caagccccgg
     2101 ctcccacccc ctcctgtacc cccacctctg ctgcattacc ctccccctgc caaggtgcca
     2161 ggcctggagc cctgcccccc acctcccttc cctcccatgg caccacccac tgctttgctg
     2221 caggaagagc ctctcttctc tcccaggttt cccttcccca ccgtccctcc tgccccagga
     2281 gtgtctccgc tgcctgctcc tgcagccttc ccacccaccc cacagtctgt ccccagccca
     2341 gcccccaccc ccttccccat agagcttcta cccttggggt attcggagcc tgcctttggg
     2401 ccttgcttct ccatgcccag aggcaagccc cccgccccat cccctagggg acagaaagcc
     2461 agccccccta ccttagcccc tgccactgcc agtcccccca ccactgcggg gagcaacaac
     2521 ccctgcctca cacagctgct cacagcagct aagccggagc aagccctgga gccaccactt
     2581 gtatccagca ccctcctccg gtccccaggg tccccgcagg agacagtccc tgaattcccc
     2641 tgcacattcc ttcccccgac cccggcccct acaccgcccc ggccacctcc aggcccggcc
     2701 acattggccc cttccaggcc cctgcttgtc cccaaagcgg agcggctctc acccccagcg
     2761 cccagcggca gtgaacggcg gctgtcaggg gacctcagct ccatgccagg ccctgggact
     2821 ctgagcgtcc gtgtctctcc cccgcaaccc atcctcagcc ggggccgtcc agacagcaac
     2881 aagaccgaga accggcgtat cacacacatc tccgcggagc agaagcggcg cttcaacatc
     2941 aagctggggt ttgacaccct tcatgggctc gtgagcacac tcagtgccca gcccagcctc
     3001 aaggtgagca aagctaccac gctgcagaag acagctgagt acatccttat gctacagcag
     3061 gagcgtgcgg gcttgcagga ggaggcccag cagctgcggg atgagattga ggagctcaat
     3121 gccgccatta acctgtgcca gcagcagctg cccgccacag gggtacccat cacacaccag
     3181 cgttttgacc agatgcgaga catgtttgat gactacgtcc gaacccgtac gctgcacaac
     3241 tggaagttct gggtgttcag catcctcatc cggcctctgt ttgagtcctt caacgggatg
     3301 gtgtccacgg caagtgtgca caccctccgc cagacctcac tggcctggct ggaccagtac
     3361 tgctctctgc ccgctctccg gccaactgtc ctgaactccc tacgccagct gggcacatct
     3421 accagtatcc tgaccgaccc gggccgcatc cctgagcaag ccacacgggc agtcacagag
     3481 ggcacccttg gcaaaccttt atagtctaga gggccctatt ctatagtgtc acctaaatgc
     3541 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3601 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3661 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3721 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3781 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3961 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4021 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4141 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4201 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4261 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg
     4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4381 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4441 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4501 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4561 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4621 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4681 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4741 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4801 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4861 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4921 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4981 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5041 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5101 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5161 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5221 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5281 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5341 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5401 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5461 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5521 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5581 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5761 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5941 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6001 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6121 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6181 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6781 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6841 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6901 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6961 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7021 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7081 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7141 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7201 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7261 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7321 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7381 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7441 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7501 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7561 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7621 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7681 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7741 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7801 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7861 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7921 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
