LOCUS       V030329                 7205 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030329
VERSION     V030329
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             941..2695
                     /label="FAM151A(NM_176782)"
                     /note="FAM151A(NM_176782)"
                     /gene="FAM151A"
     CDS             2702..2767
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2805..3029
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3075..3503
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3517..3846
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3913..4707
                     /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    4881..5002
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5051..5067)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5075..5091
                     /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(5099..5129)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5144..5165
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5453..6038)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6209..7069)
                     /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(7070..7174)
                     /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 atccgccacc atggtctgca gggagcagtt
      961 atcaaagaat caggtcaagt gggtgtttgc cggcattacc tgtgtgtctg tggtggtcat
     1021 tgccgcaata gtccttgcca tcaccctgcg gcggccaggc tgtgagctgg aggcctgcag
     1081 ccctgatgcc gacatgctgg actacctgct gagcctgggc cagatcagcc ggcgagatgc
     1141 cttggaggtc acctggtacc acgcagccaa cagcaagaaa gccatgacag ctgccctgaa
     1201 cagcaacatc acagtcctgg aggctgacgt caatgtagaa gggctcggca cagccaatga
     1261 gacaggagtt cccatcatgg cacacccccc cactatctac agtgacaaca cactggagca
     1321 gtggctggac gctgtgctgg gctcttccca aaagggcatc aaactggact tcaagaacat
     1381 caaggcagtg ggcccctccc tggacctcct gcggcagctg acagaggaag gcaaagtccg
     1441 gcggcccata tggatcaacg ctgacatctt aaagggcccc aacatgctca tctcaactga
     1501 ggtcaatgcc acacagttcc tggccctggt ccaggagaag tatcccaagg ctaccctatc
     1561 tccaggctgg accaccttct acatgtccac gtccccaaac aggacgtaca cccaagccat
     1621 ggtggagaag atgcacgagc tggtgggagg agtgccccag agggtcacct tccctgtacg
     1681 gtcttccatg gtgcgggctg cctggcccca cttcagctgg ctgctgagcc aatctgagag
     1741 gtacagcctg acgctgtggc aggctgcctc ggaccccatg tcggtggaag atctgctcta
     1801 cgtccgggat aacactgctg tccaccaagt ctactatgac atctttgagc ctctcctgtc
     1861 acagttcaag cagctggcct tgaatgccac acggaaacca atgtactaca cgggaggcag
     1921 cctgatccct cttctccagc tgcctgggga tgacggtctg aatgtggagt ggctggttcc
     1981 tgacgtccag ggcagcggta aaacagcaac aatgaccctc ccagacacag aaggcatgat
     2041 cctgctgaac actggcctcg agggaactgt ggctgaaaac cccgtgccca ttgttcatac
     2101 tccaagtggc aacatcctga cgctggagtc ctgcctgcag cagctggcca cacatcccgg
     2161 acactggggc atccatttgc aaatagcgga gcccgcagcc ctccggccat ccctggcctt
     2221 gctggcacgc ctctccagcc ttggcctctt gcattggcct gtgtgggttg gggccaaaat
     2281 ctcccacggg agtttttcgg tccccggcca tgtggctggc agagagctgc ttacagctgt
     2341 ggctgaggtc ttcccccacg tgactgtggc accaggctgg cctgaggagg tgctgggcag
     2401 tggctacagg gaacagctgc tcacagatat gctagagttg tgccaggggc tctggcaacc
     2461 tgtgtccttc cagatgcagg ccatgctgct gggccacagc acagctggag ccataggcag
     2521 gctgctggca tcctcccccc gggccaccgt cacagtggag cacaacccag ctgggggcga
     2581 ctatgcctct gtgaggacag cattgctggc agctagggct gtggacagga cccgagtcta
     2641 ctacaggcta ccccagggct accacaagga cttgctggct catgttggta gaaacctcga
     2701 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2761 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2821 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2881 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2941 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3001 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3061 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3121 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3181 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3241 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3301 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3361 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3421 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3481 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3781 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3841 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3901 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3961 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4021 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4081 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4141 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4201 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4261 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4321 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4381 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4441 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4501 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4561 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4621 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4681 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4741 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4801 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4861 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4981 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7201 acgtc
//
