LOCUS       V028558                 7638 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028558
VERSION     V028558
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2318
                     /label="Ceacam1(NM_001039185)"
                     /note="Ceacam1(NM_001039185)"
                     /gene="Ceacam1"
     CDS             2325..2390
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2414..3002
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3068..3116
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3318..3746
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3760..4089
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4156..4950
                     /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    5126..5247
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5296..5312)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5320..5336
                     /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(5344..5374)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5389..5410
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5698..6286)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6457..7317)
                     /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(7318..7422)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga gctggcctca gcacatctcc
      781 acaaagggca ggttccctgg ggaggactac tgctcacagc ctcactttta gcctcctgga
      841 gccctgccac cactgctgaa gtcaccattg aggctgtgcc gccccaggtt gctgaagaca
      901 acaatgttct tctacttgtt cacaatctgc ccctggcgct tggagccttt gcctggtaca
      961 agggaaacac tacggctata gacaaagaaa ttgcacgatt tgtaccaaat agtaatatga
     1021 atttcacggg gcaagcatac agcggcagag agataatata cagcaatgga tccctgctct
     1081 tccaaatgat caccatgaag gatatgggag tctacacact agatatgaca gatgaaaact
     1141 atcgtcgtac tcaggcgact gtgcgatttc atgtacaccc catattatta aagcccaaca
     1201 tcacaagcaa caactccaat cccgtggagg gtgacgactc cgtatcatta acctgtgact
     1261 cttacactga ccctgataat ataaactacc tgtggagcag aaatggtgaa agcctttcag
     1321 aaggtgacag gctgaagctg tctgagggca acaggactct cactttactc aatgtcacga
     1381 ggaatgacac aggaccctat gtgtgtgaaa cccggaatcc agtgagtgtc aaccgaagtg
     1441 acccattcag cctgaacatt atctatggtc cggacacccc gattatatcc ccctcagata
     1501 tttatttgca tccagggtca aacctcaacc tctcctgcca tgcagcctct aacccacctg
     1561 cacagtactt ttggcttatc aatgagaagc cccatgcatc ctcccaagag ctctttatcc
     1621 ccaacatcac tactaataat agcggaacct atacctgctt cgtcaataac tctgtcactg
     1681 gcctcagtag gaccacagtc aagaacatta cagtccttga gccagtgact cagcccttcc
     1741 tccaagtcac caacaccaca gtcaaagaac tagactctgt gaccctgacc tgcttgtcga
     1801 atgacattgg agccaacatc cagtggctct tcaatagcca gagtcttcag ctcacagaga
     1861 gaatgacact ctcccagaac aacagcatcc tcagaataga ccctattaag agggaagatg
     1921 ccggcgagta tcagtgtgaa atctcgaatc cagtcagcgt caggaggagc aactcaatca
     1981 agctggacat aatatttgac ccaacacaag gaggcctctc agatggcgcc attgctggca
     2041 tcgtgattgg agttgtggct ggggtggctc taatagcagg gctggcatat ttcctctatt
     2101 ccaggaagtc tggcggggga agtgaccagc gagatctcac agagcacaaa ccctcagcct
     2161 ccaaccacaa tctggctcct tctgacaact ctcctaacaa ggtggatgac gtcgcataca
     2221 ctgtcctgaa cttcaattcc cagcaaccca accggccaac ttcagcccct tcttctccaa
     2281 gagccacaga aacagtttat tcagaagtaa aaaagaaggg atccgactac aaagaccatg
     2341 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2401 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2461 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2521 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2581 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2641 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2701 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2761 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2821 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2881 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2941 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3001 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3061 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3121 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3181 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3241 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     3301 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3361 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3421 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3481 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3541 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3601 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3661 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3721 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3961 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4021 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4081 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4141 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4201 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4261 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4321 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4381 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4441 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4501 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4561 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4621 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4681 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4741 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4801 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4861 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4921 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4981 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5041 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5101 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5161 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5221 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5281 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5341 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5401 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5881 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6121 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7441 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7501 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7561 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7621 atgaagaatc tgcttagg
//
