LOCUS       V022075                 8255 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022075
VERSION     V022075
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..3787
                     /label="MMRN2(NM_024756)"
                     /note="MMRN2(NM_024756)"
                     /gene="MMRN2"
     CDS             3794..3817
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    3855..4079
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4125..4553
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4567..4896
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4963..5757
                     /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    5931..6052
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6101..6117)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6125..6141
                     /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(6149..6179)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6194..6215
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6503..7088)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7259..8119)
                     /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(8120..8224)
                     /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 atgatcctga gcttgctgtt
      961 cagccttggg ggccccctgg gctgggggct gctgggggca tgggcccagg cttccagtac
     1021 tagcctctct gatctgcaga gctccaggac acctggggtc tggaaggcag aggctgagga
     1081 caccggcaag gaccccgtag gacgtaactg gtgcccctac ccaatgtcca agctggtcac
     1141 cttactagct ctttgcaaaa cagagaaatt cctcatccac tcgcagcagc cgtgtccgca
     1201 gggagctcca gactgccaga aagtcaaagt catgtaccgc atggcccaca agccagtgta
     1261 ccaggtcaag cagaaggtgc tgacctcttt ggcctggagg tgctgccctg gctacacggg
     1321 ccccaactgc gagcaccacg attccatggc aatccctgag cctgcagatc ctggtgacag
     1381 ccaccaggaa cctcaggatg gaccagtcag cttcaaacct ggccaccttg ctgcagtgat
     1441 caatgaggtt gaggtgcaac aggaacagca ggaacatctg ctgggagatc tccagaatga
     1501 tgtgcaccgg gtggcagaca gcctgccagg cctgtggaaa gccctgcctg gtaacctcac
     1561 agctgcagtg atggaagcaa atcaaacagg gcacgagttc cctgatagat ccttggagca
     1621 ggtgctgcta ccccacgtgg acaccttcct acaagtgcat ttcagcccca tctggaggag
     1681 ctttaaccaa agcctgcaca gccttaccca ggccataaga aacctgtctc ttgacgtgga
     1741 ggccaaccgc caggccatct ccagagtcca ggacagtgcc gtggccaggg ctgacttcca
     1801 ggagcttggt gccaaatttg aggccaaggt ccaggagaac actcagagag tgggtcagct
     1861 gcgacaggac gtggaggacc gcctgcacgc ccagcacttt accctgcacc gctcgatctc
     1921 agagctccaa gccgatgtgg acaccaaatt gaagaggctg cacaaggctc aggaggcccc
     1981 agggaccaat ggcagtctgg tgttggcaac gcctggggct ggggcaaggc ctgagccgga
     2041 cagcctgcag gccaggctgg gccagctgca gaggaacctc tcagagctgc acatgaccac
     2101 ggcccgcagg gaggaggagt tgcagtacac cctggaggac atgagggcca ccctgacccg
     2161 gcacgtggat gagatcaagg aactgtactc cgaatcggac gagactttcg atcagattag
     2221 caaggtggag cggcaggtgg aggagctgca ggtgaaccac acggcgctcc gtgagctgcg
     2281 cgtgatcctg atggagaagt ctctgatcat ggaggagaac aaggaggagg tggagcggca
     2341 gctcctggag ctcaacctca cgctgcagca cctgcagggt ggccatgccg acctcatcaa
     2401 gtacgtgaag gactgcaatt gccagaagct ctatttagac ctggacgtca tccgggaggg
     2461 ccagagggac gccacgcgtg ccctggagga gacccaggtg agcctggacg agcggcggca
     2521 gctggacggc tcctccctgc aggccctgca gaacgccgtg gacgccgtgt cgctggccgt
     2581 ggacgcgcac aaagcggagg gcgagcgggc gcgggcggcc acgtcgcggc tccggagcca
     2641 agtgcaggcg ctggatgacg aggtgggcgc gctgaaggcg gccgcggccg aggcccgcca
     2701 cgaggtgcgc cagctgcaca gcgccttcgc cgccctgctg gaggacgcgc tgcggcacga
     2761 ggcggtgctg gccgcgctct tcggggagga ggtgctggag gagatgtctg agcagacgcc
     2821 gggaccgctg cccctgagct acgagcagat ccgcgtggcc ctgcaggacg ccgctagcgg
     2881 gctgcaggag caggcgctcg gctgggacga gctggccgcc cgagtgacgg ccctggagca
     2941 ggcctcggag cccccgcggc cggcagagca cctggagccc agccacgacg cgggccgcga
     3001 ggaggccgcc accaccgccc tggccgggct ggcgcgggag ctccagagcc tgagcaacga
     3061 cgtcaagaat gtcgggcggt gctgcgaggc cgaggccggg gccggggccg cctccctcaa
     3121 cgcctccctt cacggcctcc acaacgcact cttcgccact cagcgcagct tggagcagca
     3181 ccagcggctc ttccacagcc tctttgggaa cttccaaggg ctcatggaag ccaacgtcag
     3241 cctggacctg gggaagctgc agaccatgct gagcaggaaa gggaagaagc agcagaaaga
     3301 cctggaagct ccccggaaga gggacaagaa ggaagcggag cctttggtgg acatacgggt
     3361 cacagggcct gtgccaggtg ccttgggcgc ggcgctctgg gaggcaggat cccctgtggc
     3421 cttctatgcc agcttttcag aagggacggc tgccctgcag acagtgaagt tcaacaccac
     3481 atacatcaac attggcagca gctacttccc tgaacatggc tacttccgag cccctgagcg
     3541 tggtgtctac ctgtttgcag tgagcgttga atttggccca gggccaggca ccgggcagct
     3601 ggtgtttgga ggtcaccatc ggactccagt ctgtaccact gggcagggga gtggaagcac
     3661 agcaacggtc tttgccatgg ctgagctgca gaagggtgag cgagtatggt ttgagttaac
     3721 ccagggatca ataacaaaga gaagcctgtc gggcactgca tttgggggct tcctgatgtt
     3781 taagaccctc gaggattaca aggatgacga cgataagtag tgagggcccg tttaaacccg
     3841 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3901 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3961 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4021 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4081 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4561 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4801 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4861 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4921 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4981 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5041 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5101 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5161 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5221 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5281 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5341 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5401 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5461 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5521 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5581 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5641 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5701 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5761 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5821 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5881 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6061 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7021 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7081 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7141 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7201 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7261 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7321 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7381 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7441 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7501 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7561 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7621 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7681 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7741 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7801 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7861 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7921 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7981 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8041 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8101 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8161 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8221 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
