LOCUS       V041037                 7542 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041037
VERSION     V041037
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..2222
                     /label="Fut11(NM_028428)"
                     /note="Fut11(NM_028428)"
                     /gene="Fut11"
     CDS             2229..2294
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2318..2906
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2972..3020
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3222..3650
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3664..3993
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4060..4854
                     /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    5030..5151
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5200..5216)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5224..5240
                     /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(5248..5278)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5293..5314
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5602..6190)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6361..7221)
                     /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(7222..7326)
                     /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 ccaccatggc tgctcgctgt accgaggcgg
      781 tgctggccgc cctgggggtg ctcagtgtct gttcggccag cagctccggg tccgaggctt
      841 ccggggaggc agagagggag gagccgtggg atggcgcggt tttccggcct cccgccgccc
      901 taggcgccgt ggggatagcg cgcgggcccg ggtccccgcc tccagggaac agagaggcgg
      961 tggacctacc agtgttgctg tggtggagcc caggtctgtt tccgcacttc ccaggtgact
     1021 cggagcgcat acagtgtgcg cacggcgcgt gcgtggcgtc ccgggatcga cgggcgcgcg
     1081 ccgacccgcg gacgcgcgcg ttgctctttt acggcacgga cttccgcgcg gctgatgcgc
     1141 ccctgccgcg cctggcgcac cagagctggg cgctcctgca cgaggagtcg ccgctcaata
     1201 acttcctact gagccacggc ccgggcatcc gtctcttcaa tctcaccgcc actttcagcc
     1261 gccactcgga ttatccgctg ccgctgcagt ggctgcccgg ggccgcctat ttgcgccggc
     1321 ccgcgcctcc gccccgggag cgcgccgagt ggcgtcgccg cggatacgcg ccgttgctct
     1381 atttgcagtc ccattgcgat gtgccttccg accgggaccg atacgtacgc gagctcatgc
     1441 gctacatccc ggtggattcc tatggcaagt gcctgcagaa ccgcgagccg cccaccgtgc
     1501 gattacagga cacagccacg gccaccaccg aggatccaga actcatggcc tttttgtccc
     1561 ggtataagtt ccacttagct ctggagaatg ccatctgcaa tgattacatg acagaaaaac
     1621 tgtggcgccc catgcatctg ggtgctgtgc ccgtgtatcg tggatcaccc tctgtgaggg
     1681 actggatgcc caataatcac tctgtcatcc taattgatga ctttgagtcc cctcaaaagt
     1741 tggcagagtt tattgacttt ctggacaaaa atgatgacga gtatatgaaa tacctggcat
     1801 acaagcagcc tggaggcatc accaaccagt ttcttctgga taacctggag cacagggagt
     1861 ggggagtcaa cgatcccatg ttgcctaatt acctcaatgg ctttgagtgt ttcgtctgtg
     1921 accatgaact ggctcggctg aatgccgaga aagcacatgc atcctctcat ggggacattc
     1981 ctgtccccga gcctcgcatt gcccagtcgt cgcacatgaa ctgtccagtg cctacccctg
     2041 gctttgggaa agtcgaagag attcccgaga atgacagctg gaaggaaatg tggctacaag
     2101 attattggca aggtctctat cagggggaag ctctcactgc catgatccac aacaatgaga
     2161 cacagcagag gaaattttgg gattatgtac atgagatctt catgaaacgg aataaaaatc
     2221 tcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2281 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     2341 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2401 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2461 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2521 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2581 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2641 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2701 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     2761 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2821 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2881 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     2941 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3001 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3061 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3121 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3181 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3241 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3301 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3361 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3421 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3481 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3541 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3601 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3901 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3961 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4021 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4081 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4141 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4201 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4261 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4321 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4381 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4441 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4501 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4561 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4621 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4681 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4741 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4801 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4861 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5041 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5101 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5161 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6001 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7321 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7381 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7441 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7501 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
