LOCUS       V024259                 6814 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024259
VERSION     V024259
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"
     CDS             964..2346
                     /label="NEU3(NM_001367860)"
                     /note="NEU3(NM_001367860)"
                     /gene="NEU3"
     CDS             2353..2376
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2414..2638
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2684..3112
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3126..3455
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3522..4316
                     /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    4490..4611
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4660..4676)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4684..4700
                     /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(4708..4738)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4753..4774
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5062..5647)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5818..6678)
                     /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(6679..6783)
                     /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 ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgagac ctgcggacct gcccccgcgc cccatggaag aatccccggc gtccagctct
     1021 gccccgacag agacggagga gccggggtcc agtgcagagg tcatggaaga agtgacaaca
     1081 tgctccttca acagccctct gttccggcag gaagatgaca gagggattac ctaccggatc
     1141 ccagccctgc tctacatacc ccccacccac accttcctgg cctttgcaga gaagcgttct
     1201 acgaggagag atgaggatgc tctccacctg gtgctgaggc gagggttgag gattgggcag
     1261 ttggtacagt gggggcccct gaagccactg atggaagcca cactaccggg gcatcggacc
     1321 atgaacccct gtcctgtatg ggagcagaag agtggttgtg tgttcctgtt cttcatctgt
     1381 gtgcggggcc atgtcacaga gcgtcaacag attgtgtcag gcaggaatgc tgcccgcctt
     1441 tgcttcatct acagtcagga tgctggatgt tcatggagtg aggtgaggga cttgactgag
     1501 gaggtcattg gctcagagct gaagcactgg gccacatttg ctgtgggccc aggtcatggc
     1561 atccagctgc agtcagggag actggtcatc cctgcgtata cctactacat cccttcctgg
     1621 ttcttttgct tccagctacc atgtaaaacc aggcctcatt ctctgatgat ctacagtgat
     1681 gacctagggg tcacatggca ccatggtaga ctcattaggc ccatggttac agtagaatgt
     1741 gaagtggcag aggtgactgg gagggctggc caccctgtgc tatattgcag tgcccggaca
     1801 ccaaacaggt gccgggcaga ggcgctcagc actgaccatg gtgaaggctt tcagagactg
     1861 gccctgagtc gacagctctg tgagccccca catggttgcc aagggagtgt ggtaagtttc
     1921 cggcccctgg agatcccaca taggtgccag gactctagca gcaaagatgc acccaccatt
     1981 cagcagagct ctccaggcag ttcactgagg ctggaggagg aagctggaac accgtcagaa
     2041 tcatggctct tgtactcaca cccaaccagt aggaaacaga gggttgacct aggtatctat
     2101 ctcaaccaga cccccttgga ggctgcctgc tggtcccgcc cctggatctt gcactgtggg
     2161 ccctgtggct actctgatct ggctgctctg gaggaggagg gcttgtttgg gtgtttgttt
     2221 gaatgtggga ccaagcaaga gtgtgagcag attgccttcc gcctgtttac acaccgggag
     2281 atcctgagtc acctgcaggg ggactgcacc agccctggta ggaacccaag ccaattcaaa
     2341 agcaatctcg aggattacaa ggatgacgac gataagtagt gagggcccgt ttaaacccgc
     2401 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2461 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2521 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2581 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2641 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3361 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3481 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3541 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3601 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3661 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3721 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3781 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3841 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3901 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3961 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4021 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4081 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4141 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4201 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4261 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4321 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4381 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4441 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4501 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4561 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4621 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4681 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4741 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4801 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5581 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5641 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5701 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5761 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5821 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5881 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5941 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6001 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6061 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6121 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6181 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6241 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6301 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6361 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6421 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6481 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6541 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6601 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6661 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6721 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6781 gcgcacattt ccccgaaaag tgccacctga cgtc
//
