LOCUS       V023824                 7547 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023824
VERSION     V023824
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..3037
                     /label="MASP1(NM_001879)"
                     /note="MASP1(NM_001879)"
                     /gene="MASP1"
     CDS             3044..3109
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3147..3371
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3417..3845
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3859..4188
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4255..5049
                     /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    5223..5344
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5393..5409)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5417..5433
                     /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(5441..5471)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5486..5507
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5795..6380)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6551..7411)
                     /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(7412..7516)
                     /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 atgaggtggc tgcttctcta
      961 ttatgctctg tgcttctccc tgtcaaaggc ttcagcccac accgtggagc taaacaatat
     1021 gtttggccag atccagtcgc ctggttatcc agactcctat cccagtgatt cagaggtgac
     1081 ttggaatatc actgtcccag atgggtttcg gatcaagctt tacttcatgc acttcaactt
     1141 ggaatcctcc tacctttgtg aatatgacta tgtgaaggta gaaactgagg accaggtgct
     1201 ggcaaccttc tgtggcaggg agaccacaga cacagagcag actcccggcc aggaggtggt
     1261 cctctcccct ggctccttca tgtccatcac tttccggtca gatttctcca atgaggagcg
     1321 tttcacaggc tttgatgccc actacatggc tgtggatgtg gacgagtgca aggagaggga
     1381 ggacgaggag ctgtcctgtg accactactg ccacaactac attggcggct actactgctc
     1441 ctgccgcttc ggctacatcc tccacacaga caacaggacc tgccgagtgg agtgcagtga
     1501 caacctcttc actcaaagga ctggggtgat caccagccct gacttcccaa acccttaccc
     1561 caagagctct gaatgcctgt ataccatcga gctggaggag ggtttcatgg tcaacctgca
     1621 gtttgaggac atatttgaca ttgaggacca tcctgaggtg ccctgcccct atgactacat
     1681 caagatcaaa gttggtccaa aagttttggg gcctttctgt ggagagaaag ccccagaacc
     1741 catcagcacc cagagccaca gtgtcctgat cctgttccat agtgacaact cgggagagaa
     1801 ccggggctgg aggctctcat acagggctgc aggaaatgag tgcccagagc tacagcctcc
     1861 tgtccatggg aaaatcgagc cctcccaagc caagtatttc ttcaaagacc aagtgctcgt
     1921 cagctgtgac acaggctaca aagtgctgaa ggataatgtg gagatggaca cattccagat
     1981 tgagtgtctg aaggatggga cgtggagtaa caagattccc acctgtaaaa ttgtagactg
     2041 tagagcccca ggagagctgg aacacgggct gatcaccttc tctacaagga acaacctcac
     2101 cacatacaag tctgagatca aatactcctg tcaggagccc tattacaaga tgctcaacaa
     2161 taacacaggt atatatacct gttctgccca aggagtctgg atgaataaag tattggggag
     2221 aagcctaccc acctgccttc cagtgtgtgg gctccccaag ttctcccgga agctgatggc
     2281 caggatcttc aatggacgcc cagcccagaa aggcaccact ccctggattg ccatgctgtc
     2341 acacctgaat gggcagccct tctgcggagg ctcccttcta ggctccagct ggatcgtgac
     2401 cgccgcacac tgcctccacc agtcactcga tccggaagat ccgaccctac gtgattcaga
     2461 cttgctcagc ccttctgact tcaaaatcat cctgggcaag cattggaggc tccggtcaga
     2521 tgaaaatgaa cagcatctcg gcgtcaaaca caccactctc cacccccagt atgatcccaa
     2581 cacattcgag aatgacgtgg ctctggtgga gctgttggag agcccagtgc tgaatgcctt
     2641 cgtgatgccc atctgtctgc ctgagggacc ccagcaggaa ggagccatgg tcatcgtcag
     2701 cggctggggg aagcagttct tgcaaaggtt cccagagacc ctgatggaga ttgaaatccc
     2761 gattgttgac cacagcacct gccagaaggc ttatgccccg ctgaagaaga aagtgaccag
     2821 ggacatgatc tgtgctgggg agaaggaagg gggaaaggac gcctgtgcgg gtgactctgg
     2881 aggccccatg gtgaccctga atagagaaag aggccagtgg tacctggtgg gcactgtgtc
     2941 ctggggtgat gactgtggga agaaggaccg ctacggagta tactcttaca tccaccacaa
     3001 caaggactgg atccagaggg tcaccggagt gaggaacctc gaggactaca aagaccatga
     3061 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     3121 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3181 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3241 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3301 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3361 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3421 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3481 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3541 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3601 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3661 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3721 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3781 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3841 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4201 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4261 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4321 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4381 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4441 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4501 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4561 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4621 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4681 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4741 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4801 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4861 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4921 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4981 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5041 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5101 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5221 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5341 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6301 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
