LOCUS       V029793                 6992 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029793
VERSION     V029793
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      935..944
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             944..1033
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1040..2482
                     /label="Lbp(NM_008489)"
                     /note="Lbp(NM_008489)"
                     /gene="Lbp"
     CDS             2489..2554
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2592..2816
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2862..3290
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3304..3633
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3700..4494
                     /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    4668..4789
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4838..4854)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4862..4878
                     /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(4886..4916)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4931..4952
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5240..5825)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5996..6856)
                     /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(6857..6961)
                     /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 ccgagctcga attcgccacc atggaacaaa aactaatatc
      961 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc
     1021 cgaggaggac ttaggatcca tgaaggctgg gactggacct ctgctctcta cattgctggg
     1081 gctgctgttc ctgtccatac agggaactgg cggtgtcaac cccggtgtgg tggccaggat
     1141 caccgacaag ggcctggcct acgcggccaa ggaggggctg gtggctctgc agagagagct
     1201 gtacaagatc acactaccgg acttcagcgg ggacttcaag atcaaggccg tgggtcgtgg
     1261 gcagtacgag tttcatagcc tggagattca gaactgtgag ctgcgtggct cctctctgaa
     1321 gctgctccca ggccagggcc tgagtctcgc catctctgac tcttccatcg gtgtccgagg
     1381 caaatggaag gtgcgcaaat ccttcctgaa acttcatggc tcctttgacc tggatgtcaa
     1441 aggtgtcacc atttcagtgg acctcctcct gggcatggat ccctcagggc ggcccacagt
     1501 ctctgcctct gggtgcagca gccgcatttg tgatttggat gtgcacatat caggaaatgt
     1561 ggggtggctg ctgaatctct tccacaacca gatcgagtcg aagctccaga aagtactgga
     1621 gaataaggtt tgtgagatga tccagaagtc tgtgacctct gacctgcagc cttatctcca
     1681 gactctgcca gtcacagcag agatcgacaa tgtcctgggc attgactaca gtttggtggc
     1741 ggctccccaa gcgaaggccc aggtgctgga tgtcatgttt aagggtgaaa tttttaatcg
     1801 gaatcaccgc tctccagttg ctacccccac tccgaccatg agcctacctg aggacagtaa
     1861 gcaaatggtc tactttgcca tctcagatta tgccttcaac atagccagcc gggtttatca
     1921 ccaggctggg tacctgaatt tttccatcac agatgacatg ttaccacatg actccggcat
     1981 ccggttgaac accaaggcct tccgtccctt cactccccag atatacaaaa agtatcccga
     2041 catgaaattg gagctccttg gaacagtggt ctctgccccg attctgaatg tcagtcctgg
     2101 gaatctgtcc ttggccccac agatggagat cgaaggcttt gtgatcctgc ccacctccgc
     2161 ccgggaacct gtcttccggc ttggcgtggt cactaatgta tttgcctcat taactttcaa
     2221 caacagcaag gttaccggga tgctgcatcc agacaaggca caagtgagac tgatcgaatc
     2281 caaagtcggc atgttcaatg tgaacctgtt ccaggcattc cttaactact accttctcaa
     2341 cagcctctac cctgatgtca atgctgagct ggcccagggc ttccccctcc ctctgccaag
     2401 gcatattcag ctccacgacc tcgacttcca gatccgcaag gacttcttat acttgggtgc
     2461 caatgtccag tatatgagag tcctcgagga ctacaaagac catgacggtg attataaaga
     2521 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     2581 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2641 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2701 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2761 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2821 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3301 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3421 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3541 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3601 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3661 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3721 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3781 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3841 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3901 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3961 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4021 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4081 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4141 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4201 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4261 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4321 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4381 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4441 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4501 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4561 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4621 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4681 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4741 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4801 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5761 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5941 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6001 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6061 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6121 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6181 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6241 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6301 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6361 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6421 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6481 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6541 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6601 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6661 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6721 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6781 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6841 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6901 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6961 gcacatttcc ccgaaaagtg ccacctgacg tc
//
