LOCUS       V045811                 6472 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045811
VERSION     V045811
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      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..1992
                     /label="SUCNR1(NM_033050)"
                     /note="SUCNR1(NM_033050)"
                     /gene="SUCNR1"
     polyA_signal    2072..2296
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2342..2770
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2784..3113
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3180..3974
                     /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    4148..4269
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4318..4334)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4342..4358
                     /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(4366..4396)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4411..4432
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4720..5305)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5476..6336)
                     /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(6337..6441)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgctgggga tcatggcatg gaatgcaact
     1021 tgcaaaaact ggctggcagc agaggctgcc ctggaaaagt actacctttc cattttttat
     1081 gggattgagt tcgttgtggg agtccttgga aataccattg ttgtttacgg ctacatcttc
     1141 tctctgaaga actggaacag cagtaatatt tatctcttta acctctctgt ctctgactta
     1201 gcttttctgt gcaccctccc catgctgata aggagttatg ccaatggaaa ctggatatat
     1261 ggagacgtgc tctgcataag caaccgatat gtgcttcatg ccaacctcta taccagcatt
     1321 ctctttctca cttttatcag catagatcga tacttgataa ttaagtatcc tttccgagaa
     1381 caccttctgc aaaagaaaga gtttgctatt ttaatctcct tggccatttg ggttttagta
     1441 accttagagt tactacccat acttcccctt ataaatcctg ttataactga caatggcacc
     1501 acctgtaatg attttgcaag ttctggagac cccaactaca acctcattta cagcatgtgt
     1561 ctaacactgt tggggttcct tattcctctt tttgtgatgt gtttctttta ttacaagatt
     1621 gctctcttcc taaagcagag gaataggcag gttgctactg ctctgcccct tgaaaagcct
     1681 ctcaacttgg tcatcatggc agtggtaatc ttctctgtgc tttttacacc ctatcacgtc
     1741 atgcggaatg tgaggatcgc ttcacgcctg gggagttgga agcagtatca gtgcactcag
     1801 gtcgtcatca actcctttta cattgtgaca cggcctttgg cctttctgaa cagtgtcatc
     1861 aaccctgtct tctattttct tttgggagat cacttcaggg acatgctgat gaatcaactg
     1921 agacacaact tcaaatccct tacatccttt agcagatggg ctcatgaact cctactttca
     1981 ttcagagaaa agtgagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     2041 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2281 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2341 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2761 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2941 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3001 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3061 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3121 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3181 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3241 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3301 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3361 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3421 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3481 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3541 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3601 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3661 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3721 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3781 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3841 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3901 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3961 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4021 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4081 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4141 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4201 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4261 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     5281 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5341 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5401 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5461 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5521 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5581 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5641 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5701 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5761 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5821 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5881 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5941 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6001 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6061 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6121 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6181 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6241 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6301 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6361 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6421 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
