LOCUS       V043695                 7075 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043695
VERSION     V043695
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..2565
                     /label="Lpcat1(NM_145376)"
                     /note="Lpcat1(NM_145376)"
                     /gene="Lpcat1"
     CDS             2572..2637
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2675..2899
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2945..3373
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3387..3716
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3783..4577
                     /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    4751..4872
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4921..4937)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4945..4961
                     /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(4969..4999)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5014..5035
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5323..5908)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6079..6939)
                     /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(6940..7044)
                     /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 accatgaggc tgcggggccg cgggccgcgg gccgccccct cctccagctc tggggcaggc
     1021 gacgcgcggc gcctagcgcc cccggggcgg aaccccttcg tgcacgagct gcgactgagc
     1081 gccctgcaga aggcccaggt ggctttcatg acgttgacgc tgttccccat ccggctcctg
     1141 ttcgctgctt tcatgatgct gctggcctgg ccctttgcac tcctggcttc cctgggacct
     1201 cctgataagg agccagagca gcccctggcc ttatggagga aggtcgtgga cttcctgctc
     1261 aaggccatca tgcgcaccat gtggtttgct ggcggcttcc accgtgtagc tgtgaagggg
     1321 cggcaggccc tgcctactga ggctgctatc ctcaccttgg ctccacattc ctcctacttt
     1381 gatgccatcc cggtcaccat gaccatgtcc tccattgtga tgaaggcaga gagcagagac
     1441 atcccaatct ggggaactct gataagatac atcaggccag tgttcgtgtc ccgctctgac
     1501 caggactcgc gaaggaagac agtggaggag atcaagcgac gggcacagtc gaatggaaag
     1561 tggcctcaga taatgatttt tccagaagga acttgtacaa ataggacctg cctcattacc
     1621 ttcaaacctg gtgcgttcat tcctggagtt cctgtccagc ccgtggtgct acgctaccca
     1681 aacaaattgg acaccatcac atggacgtgg caaggacctg gagcgttgaa aatcctgtgg
     1741 ctcactctgt gccagtttca aaaccaagtg gaaattgaat ttctgcctgt gtattgccct
     1801 tctgaagagg agaagaggaa tcctgccctg tatgccagca atgtgaggcg tgtcatggcc
     1861 aaggccctgg gtgtctcagt aaccgactat acatttgagg actgccagct ggctctggca
     1921 gaaggacaac ttcgcttgcc tgctgacacc tgcctgctag agtttgccag gcttgtgagg
     1981 ggcctcggac taaaaccaga aaatcttgaa aaagatctag acaaatattc agaaagtgcg
     2041 aggatgaaga gaggagagaa aatccgcctt ccagagtttg cggcctactt ggaagttcct
     2101 gtctcagatg cactggaaga catgttctcg ctttttgacg agagtggtgg tggtgagatt
     2161 gaccttcgtg aatatgtggt cgccttgtct gtggtgtgca ggccatccca gaccttagcc
     2221 accatccagc tggcattcaa gatgtacgga tctcctgagg atggcagcat agatgaggcc
     2281 aacctgtcct gcatcctcaa gactgcactg ggtgtatcag aattaacagt gaccgacttg
     2341 ttccaggcta ttgaccaaga ggacaaggga agaatcacct ttgatgactt ctgcgggttt
     2401 gcggaaatgt accccgacta cgcagaggac tacttgtacc ctgatcagac acatttcgac
     2461 agctgtgcac agacaccccc agcaccaact cccaatggct tctgcattga cttcagccct
     2521 gagaactcag actttgggag aaagaattct tgtaagaaag cggacctcga ggactacaaa
     2581 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     2641 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2701 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2761 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2821 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2881 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2941 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3001 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3061 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3121 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3181 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3241 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3301 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3361 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3541 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3601 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3661 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3721 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3781 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3841 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3901 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3961 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4021 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4081 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4141 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4201 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4261 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4321 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4381 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4441 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4501 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4561 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4621 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4681 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4741 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4801 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4861 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4921 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4981 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5041 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5101 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5161 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
