LOCUS       V041418                 9622 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041418
VERSION     V041418
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..5112
                     /label="CEMIP2(NM_013390)"
                     /note="CEMIP2(NM_013390)"
                     /gene="CEMIP2"
     CDS             5119..5184
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5222..5446
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5492..5920
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5934..6263
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6330..7124
                     /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    7298..7419
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7468..7484)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7492..7508
                     /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(7516..7546)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7561..7582
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7870..8455)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8626..9486)
                     /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(9487..9591)
                     /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 accatgtatg ccactgattc caggggacac tcccctgctt tcctccaacc tcagaatgga
     1021 aatagtcgtc acccatctgg ctatgttcca gggaaggttg tcccattgcg tccccctcct
     1081 cctccaaaga gtcaagcttc agccaaattt acctccatca gacgagaaga ccgggcaacc
     1141 ttcgcattct cacctgaaga acagcaagcc cagagagaaa gtcaaaagca aaagagacac
     1201 aaaaatactt tcatttgttt tgctattact agtttctcat tttttattgc acttgcaatc
     1261 attttaggaa tatcctcaaa atatgctcca gatgaaaatt gcccagatca aaatcctcgt
     1321 ctcaggaatt gggatccagg acaagattct gcaaagcaag ttgttatcaa ggagggagat
     1381 atgctccgtc tgacctcaga cgccaccgtg cattctatag tcattcagga tggaggactg
     1441 cttgtatttg gggacaataa agatggatcc agaaatatta ctttgaggac tcattacatc
     1501 ctgatccagg atggtggggc gcttcatatt ggagcagaaa aatgccgcta taaatccaaa
     1561 gcgacaatta ccttgtatgg caagtcagat gaaggtgaaa gtatgccaac atttggcaaa
     1621 aagtttattg gtgtggaagc tggcgggaca ctggagttac atggggcacg gaaggcatcg
     1681 tggacgttgt tggcaaggac cctgaattcc tcaggcttgc cctttgggtc ctataccttt
     1741 gaaaaggact tttcccgggg cctcaatgtg agggtcattg accaagacac ggccaaaatt
     1801 ttggaaagtg agagatttga tacccatgaa taccgcaatg agagcaggcg gcttcaggag
     1861 tttctgagat tccaggatcc aggtcggatt gttgccatag ctgtcgggga ttcagccgct
     1921 aaaagtctct tacaaggaac catccagatg atccaggaac ggttgggaag tgaactgatc
     1981 caaggactgg gctacaggca agcttgggct ttagttggtg tcattgatgg tggaagcact
     2041 tcttgcaatg aatccgtgag aaactatgaa aatcatagca gtggcgggaa ggctcttgcc
     2101 caaagagaat tttatactgt ggatggccag aagttctctg tgacagctta tagtgaatgg
     2161 attgaaggcg tttctctttc aggattccgg gtagaggttg tagatggagt gaagctaaat
     2221 ttgctagatg atgttagtag ttggaaacct ggagaccaga ttgtggtcgc aagcacagac
     2281 tattccatgt accaagcaga ggagttcact cttcttccct gttctgaatg cagccatttt
     2341 caggtcaaag tcaaagaaac ccctcagttc ctgcacatgg gtgagatcat agacggtgta
     2401 gacatgagag ctgaggttgg aattcttacc cggaatattg tgatccaagg agaagtggag
     2461 gactcatgct acgcagaaaa tcagtgccaa ttttttgatt atgatacctt tgggggacac
     2521 attatgataa tgaaaaattt tacttcagtc catctttctt atgtggaatt gaaacacatg
     2581 ggtcagcagc agatggggcg ataccctgtt cattttcacc tgtgtggtga cgtggattat
     2641 aaaggaggat acagacatgc aacatttgtg gacggcctgt ctattcatca cagcttctca
     2701 aggtgcatca ctgtgcatgg gacaaatggc ttgctaataa aagacaccat tgggtttgac
     2761 acactaggtc attgtttctt tttggaagat ggtattgaac agaggaatac tttgttccac
     2821 aatctgggac tcctcaccaa gccgggtact ctcctgccca ccgataggaa caactccatg
     2881 tgtaccacca tgcgagataa agtgtttgga aattacattc ctgtgcctgc tactgactgt
     2941 atggctgttt caactttctg gattgctcat cccaacaata atctgattaa taatgcagct
     3001 gcaggctcac aggatgctgg aatatggtat ttattccaca aggaaccaac tggggaatcc
     3061 agtggattgc agctcttggc aaaaccagaa ctcactccat tgggtatatt ttataacaac
     3121 agggtccatt caaattttaa ggctggctta tttattgaca aaggtgtcaa aacaaccaac
     3181 tctagtgctg ctgacccaag ggaatacctc tgtttggaca atagtgcaag atttcgacct
     3241 catcaggatg caaaccccga aaaaccacgt gttgctgctc taattgacag gctcattgct
     3301 tttaaaaata atgataatgg agcttgggtc agaggaggag atattatcgt tcaaaattca
     3361 gcatttgcag ataatggaat aggactgacc tttgccagtg atggaagctt cccaagtgat
     3421 gaaggttcca gccaagaggt atctgaatct ctctttgttg gggagagcag gaattacggc
     3481 tttcagggtg gtcagaacaa gtatgtaggc actggaggaa tagaccagaa gcctcgaaca
     3541 ttacccagga acaggacgtt cccaattaga ggctttcaga tttatgatgg gcccattcat
     3601 ctcacaagga gcactttcaa aaaatatgtg ccaactccag ataggtacag cagtgcaatt
     3661 ggcttcctca tgaagaattc ctggcagata acccccagga ataatatctc cctcgtgaag
     3721 tttggtccac atgtctctct gaatgtcttt tttggaaagc ctggtccctg gtttgaagat
     3781 tgtgagatgg atggtgataa gaactccata ttccatgaca ttgatggctc tgtgacagga
     3841 tacaaggatg cttatgtggg aagaatggac aactacctga tccgccatcc aagctgtgta
     3901 aatgtgtcta agtggaatgc agtgatctgc agtgggacct atgcacaggt ctatgtacag
     3961 acatggagca ctcagaatct ttctatgacc attacacgag atgagtatcc gtccaaccct
     4021 atggtgctcc gaggtattaa tcagaaggct gcctttccac agtaccagcc tgtcgtcatg
     4081 ctggagaagg gttataccat ccactggaat gggccggcac cacggactac atttctatac
     4141 ctcgtcaact tcaacaagaa tgactggatt cgagttggcc tttgctatcc atcaaacaca
     4201 agttttcaag ttacctttgg ctatttgcag cggcagaatg gctcattatc caaaatcgaa
     4261 gaatatgagc ctgtgcattc actggaagaa ctgcaaagaa agcaatccga gaggaaattc
     4321 tattttgact ccagcacggg gttactgttt ttgtatctca aagccaaaag ccacaggcat
     4381 ggccacagtt actgttcatc tcagggatgt gaaagagtca agatccaagc agccacagac
     4441 tcaaaggaca tcagtaactg catggccaaa gcatacccac agtactacag aaagccgtca
     4501 gtggtcaagc ggatgccggc catgctcact ggactctgtc aaggctgtgg cactcggcag
     4561 gtggtgttta ctagtgatcc tcataaaagt tacctccctg tgcaattcca gtcacctgat
     4621 aaagcagaaa cccagcgtgg agacccgtct gttatttctg tcaatggcac tgactttacc
     4681 ttccgaagtg caggcgtcct cctccttgtt gtggatccgt gcagcgttcc attccgcttg
     4741 acggaaaaaa cggtttttcc tcttgctgat gtcagtcgca ttgaagagta tttaaaaaca
     4801 ggcatccctc caaggtccat tgttctgttg agcacaagag gagaaataaa gcagttaaac
     4861 atttcacact tactagtacc tctgggatta gccaaaccag ctcatcttta tgacaaaggg
     4921 agtaccatat ttttgggatt cagtggaaac tttaaaccat catggactaa gctatttacc
     4981 agtcctgctg gacagggcct tggggtgctt gaacaattca tacctttgca gctggacgaa
     5041 tatggttgtc ccagagccac cactgtccgc agaagagacc tggaactgct aaagcaagct
     5101 tcaaaagcac atctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc
     5161 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg
     5221 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5281 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5341 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5401 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     5461 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     5521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5641 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6481 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6841 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     7021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     7141 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     7201 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     7261 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     7321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7441 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7561 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8281 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8401 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8701 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8761 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8821 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8881 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8941 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9001 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9061 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9121 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9181 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9241 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9301 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9361 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9421 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9481 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9601 ccgaaaagtg ccacctgacg tc
//
