LOCUS       V018549                 7282 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018549
VERSION     V018549
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             1009..2835
                     /label="AFP(NM_001134)"
                     /note="AFP(NM_001134)"
                     /gene="AFP"
     polyA_signal    2882..3106
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3152..3580
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3594..3923
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3990..4784
                     /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    4958..5079
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5128..5144)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5152..5168
                     /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(5176..5206)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5221..5242
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5530..6115)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6286..7146)
                     /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(7147..7251)
                     /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 ggtaccgagc tcggatccat gaagtgggtg
     1021 gaatcaattt ttttaatttt cctactaaat tttactgaat ccagaacact gcatagaaat
     1081 gaatatggaa tagcttccat attggattct taccaatgta ctgcagagat aagtttagct
     1141 gacctggcta ccatattttt tgcccagttt gttcaagaag ccacttacaa ggaagtaagc
     1201 aaaatggtga aagatgcatt gactgcaatt gagaaaccca ctggagatga acagtcttca
     1261 gggtgtttag aaaaccagct acctgccttt ctggaagaac tttgccatga gaaagaaatt
     1321 ttggagaagt acggacattc agactgctgc agccaaagtg aagagggaag acataactgt
     1381 tttcttgcac acaaaaagcc cactccagca tcgatcccac ttttccaagt tccagaacct
     1441 gtcacaagct gtgaagcata tgaagaagac agggagacat tcatgaacaa attcatttat
     1501 gagatagcaa gaaggcatcc cttcctgtat gcacctacaa ttcttctttg ggctgctcgc
     1561 tatgacaaaa taattccatc ttgctgcaaa gctgaaaatg cagttgaatg cttccaaaca
     1621 aaggcagcaa cagttacaaa agaattaaga gaaagcagct tgttaaatca acatgcatgt
     1681 gcagtaatga aaaattttgg gacccgaact ttccaagcca taactgttac taaactgagt
     1741 cagaagttta ccaaagttaa ttttactgaa atccagaaac tagtcctgga tgtggcccat
     1801 gtacatgagc actgttgcag aggagatgtg ctggattgtc tgcaggatgg ggaaaaaatc
     1861 atgtcctaca tatgttctca acaagacact ctgtcaaaca aaataacaga atgctgcaaa
     1921 ctgaccacgc tggaacgtgg tcaatgtata attcatgcag aaaatgatga aaaacctgaa
     1981 ggtctatctc caaatctaaa caggttttta ggagatagag attttaacca attttcttca
     2041 ggggaaaaaa atatcttctt ggcaagtttt gttcatgaat attcaagaag acatcctcag
     2101 cttgctgtct cagtaattct aagagttgct aaaggatacc aggagttatt ggagaagtgt
     2161 ttccagactg aaaaccctct tgaatgccaa gataaaggag aagaagaatt acagaaatac
     2221 atccaggaga gccaagcatt ggcaaagcga agctgcggcc tcttccagaa actaggagaa
     2281 tattacttac aaaatgcgtt tctcgttgct tacacaaaga aagcccccca gctgacctcg
     2341 tcggagctga tggccatcac cagaaaaatg gcagccacag cagccacttg ttgccaactc
     2401 agtgaggaca aactattggc ctgtggcgag ggagcggctg acattattat cggacactta
     2461 tgtatcagac atgaaatgac tccagtaaac cctggtgttg gccagtgctg cacttcttca
     2521 tatgccaaca ggaggccatg cttcagcagc ttggtggtgg atgaaacata tgtccctcct
     2581 gcattctctg atgacaagtt cattttccat aaggatctgt gccaagctca gggtgtagcg
     2641 ctgcaaacga tgaagcaaga gtttctcatt aaccttgtga agcaaaagcc acaaataaca
     2701 gaggaacaac ttgaggctgt cattgcagat ttctcaggcc tgttggagaa atgctgccaa
     2761 ggccaggaac aggaagtctg ctttgctgaa gagggacaaa aactgatttc aaaaactcgt
     2821 gctgctttgg gagtttaact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2881 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2941 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3001 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3061 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3121 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3841 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3901 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3961 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4021 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4081 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4141 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4201 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4261 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4321 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4381 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4441 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4501 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4561 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4621 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4681 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4741 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4801 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4861 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4921 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5101 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5161 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5221 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5281 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5461 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5521 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5581 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5641 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5701 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5761 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5821 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5881 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5941 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6001 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6061 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7261 ccgaaaagtg ccacctgacg tc
//
