LOCUS       V043706                 7153 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043706
VERSION     V043706
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..2643
                     /label="Sptlc2(NM_011479)"
                     /note="Sptlc2(NM_011479)"
                     /gene="Sptlc2"
     CDS             2650..2715
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2753..2977
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3023..3451
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3465..3794
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3861..4655
                     /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    4829..4950
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4999..5015)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5023..5039
                     /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(5047..5077)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5092..5113
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5401..5986)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6157..7017)
                     /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(7018..7122)
                     /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 accatgcggc cggagcccgg aggctgctgc tgccgccgcc cgatgcgggc gaacggctgc
     1021 gtcaagaacg gggaagtgag gaacgggtac ttgaggagca gcaccgccac cgtcgcggct
     1081 gccggccaga ttcatcatgt aacagaaaat ggaggactgt acaaaagacc gtttaatgaa
     1141 gcttttgaag aaacacccat gctggttgct gtgctcacat atgtgggcta tggcgtactc
     1201 accctctttg gatatcttcg agatttcttg aggcattgga gaattgaaaa gtgccaccat
     1261 gcaacagaaa gagaagaaca aaaggacttt gtgtccttgt atcaggattt tgaaaacttc
     1321 tatacaagga acctctacat gagaatcaga gacaactgga atcgacctat ctgtagtgtg
     1381 cctggagcca aggtggatat catggagaga aaatctcatg actataactg gtcattcaag
     1441 tacacaggga atataattaa aggtgtaata aacatgggtt cctacaacta tcttggattt
     1501 gcgaggaaca ctggatcatg tcaggaagca gctgctgaag tcctcaagga gtatggagca
     1561 ggggtgtgca gcactcgtca ggaaattgga aacctggaca agcatgaaga actagagaaa
     1621 ttggtagcaa ggttcttagg tgtggaagct gctatgacct atggcatggg atttgcaaca
     1681 aattcaatga acattcctgc tcttgttggc aaaggttgcc tgattctgag tgatgagctg
     1741 aaccatgcgt cactggttct aggagccaga ctgtcaggag caaccattcg aatcttcaaa
     1801 cacaacaata tgcaaagctt agagaagctt ttaaaagatg ccattgttta tggtcagcct
     1861 cggacaagaa gaccctggaa gaaaatctta atccttgtgg aaggcatata tagtatggag
     1921 gggtctattg ttcgccttcc tgaagtgatt gctctcaaga agaaatacaa ggcatacttg
     1981 tatctggatg aggctcacag cattggggct cttggccctt cagggcgagg cgtggtagat
     2041 tactttggcc tggatcctga ggatgtagat gttatgatgg gaacattcac aaagagcttc
     2101 ggtgcttcag gaggatacat cggaggcaag aaggagctga tagactacct gcgcacacat
     2161 tctcacagtg ctgtgtatgc cacgtcgatg tcaccgcctg tgatggaaca gattatcacc
     2221 tccatgaagt gcatcatggg gcaggatggc accagtcttg gcaaagaatg tatacagcag
     2281 ttggctgaga acaccaggta tttcaggaga cgcctgaagg aaatggggtt catcatctat
     2341 ggcaatgaag actccccggt ggtgcctttg atgctctaca tgccggccaa aattggcgcc
     2401 tttggaagag agatgctgaa gcggaacatt ggtgtagttg tggtgggatt tcctgctacc
     2461 ccgatcattg agtccagagc cagattttgc ctgtcagcag ctcataccaa agaaatactt
     2521 gacactgctt tgaaggagat agatgaagtt ggggatctgc tgcagctaaa gtactctcgc
     2581 caccggctgg tgcctctact ggacaggccc tttgatgaga ctacctatga agagacagaa
     2641 gacctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     2701 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2761 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2821 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2881 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2941 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3001 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3061 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3121 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3181 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3241 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3301 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3361 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3421 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3661 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3721 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3781 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3841 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3901 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3961 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4021 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4081 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4141 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4201 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4261 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4321 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4381 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4441 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4501 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4561 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4621 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4681 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4741 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4801 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4861 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4921 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4981 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5041 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5101 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5161 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5221 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5281 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5341 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5401 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5461 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5521 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5581 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5641 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5701 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5761 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5821 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5881 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     5941 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6001 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6061 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6121 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6181 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6241 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6301 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6361 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6421 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6481 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6541 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6601 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6661 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6721 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6781 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6841 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6901 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6961 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7021 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7081 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7141 gccacctgac gtc
//
