LOCUS       V044339                 8440 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044339
VERSION     V044339
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..1698
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1735..3993
                     /label="ABCB7(NM_004299)"
                     /note="ABCB7(NM_004299)"
                     /gene="ABCB7"
     polyA_signal    4040..4264
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4310..4738
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4752..5081
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5148..5942
                     /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    6116..6237
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6286..6302)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6310..6326
                     /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(6334..6364)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6379..6400
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6688..7273)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7444..8304)
                     /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(8305..8409)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggtgagca agggcgagga ggataacatg
     1021 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1081 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1141 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     1201 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     1261 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1321 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     1381 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1441 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     1501 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     1561 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     1621 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     1681 atggacgagc tgtacaaggg aggtggaggt agtggaggag gcggatcagg atccatggcg
     1741 ctgctcgcga tgcattcttg gcgctgggcg gccgcggcgg ctgctttcga aaagcgccgg
     1801 cactccgcga ttctgatccg gcctttagtc tctgttagcg gctcaggtcc gcagtggagg
     1861 ccacatcaac tcggcgcctt gggaaccgct cgagcctacc agcagattcc agagtcatta
     1921 aaaagtatca catggcagag attgggaaaa ggcaattcag gacagttctt agatgctgca
     1981 aaggctctcc aggtatggcc actgatagaa aagaggacat gttggcatgg tcatgcagga
     2041 ggaggactcc acacagaccc aaaagaaggg ttaaaagatg ttgatactcg gaaaatcata
     2101 aaagcaatgc tttcttatgt gtggcccaaa gacaggccag atctacgagc tagagttgcc
     2161 atttcgctgg gatttttggg tggtgcaaag gccatgaata ttgtggttcc cttcatgttt
     2221 aaatatgctg tagacagcct caaccagatg tcgggaaaca tgctgaacct gagtgatgca
     2281 ccaaatacag ttgcaaccat ggcaacagca gttctgattg gctatggtgt atcaagagct
     2341 ggagctgctt tttttaacga agttcgaaat gcagtatttg gcaaggtagc ccagaattca
     2401 atccgaagaa tagccaaaaa tgtctttctc catcttcaca acctggatct gggttttcac
     2461 ctgagcagac agacgggagc tttatctaag gctattgaca gaggaacaag gggtatcagt
     2521 tttgtcctga gtgctttggt atttaatctt cttcccatca tgtttgaagt gatgcttgtc
     2581 agtggtgttt tgtattacaa atgcggtgcc cagtttgctt tggtaaccct tggaacactt
     2641 ggtacataca cagcattcac agttgcagtc acacggtgga gaactagatt tagaatagaa
     2701 atgaacaaag cagataatga tgcaggtaat gctgctatag actcactgct gaattatgaa
     2761 actgtgaagt attttaataa tgaaagatat gaagcacaga gatatgatgg atttttgaag
     2821 acgtatgaga ctgcttcatt gaaaagtacc tctactctgg ctatgctgaa ctttggtcaa
     2881 agtgctattt tcagtgtcgg tttaacagct ataatggtgc tcgccagtca gggaattgtg
     2941 gcaggtaccc ttactgttgg agatctagta atggtgaatg gactgctttt tcagctttca
     3001 ttacccctga actttctggg aactgtatat agagagacta gacaagcact catagatatg
     3061 aacaccttgt ttactctact caaggtagac acccaaatta aagacaaagt gatggcatct
     3121 ccccttcaga tcacaccaca gacagctacc gtggcctttg ataatgtgca ttttgaatac
     3181 attgagggcc agaaagtcct tagtggaata tcctttgaag tccctgcagg aaagaaagtg
     3241 gccattgtag gaggtagtgg gtcagggaaa agcacaatag tgaggctatt atttcgcttc
     3301 tatgagcctc aaaagggtag catttatctt gctggtcaaa atatacaaga tgtgagcctg
     3361 gaaagccttc ggagggcagt gggagtggta cctcaggatg ctgtcctctt ccataatact
     3421 atttattaca acctcttata tggaaacatc agtgcttcac ctgaggaagt gtatgcagtg
     3481 gcaaaattag ctggacttca tgatgcaatt cttcgaatgc cacatggata tgacacccaa
     3541 gtaggggaac gaggactcaa gctttcagga ggagaaaagc aaagagtagc aattgcaaga
     3601 gccattttga aggacccccc agtcatactc tatgatgaag ctacttcatc gttagattcg
     3661 attactgaag agactattct tggtgccatg aaggatgtgg tcaaacacag aacttctatt
     3721 ttcattgcac acagattgtc aacagtggtt gatgcagatg aaatcattgt cttggatcag
     3781 ggtaaggtag ccgaacgtgg tacccaccat ggtttgcttg ctaaccctca tagtatctat
     3841 tcagaaatgt ggcatacaca gagcagccgt gtgcagaacc atgataaccc caaatgggaa
     3901 gcaaagaaag aaaatatatc caaagaggag gaaagaaaga aactacaaga agaaattgtc
     3961 aatagtgtga aaggctgtgg aaactgttcg tgctaactcg agtctagagg gcccgtttaa
     4021 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4081 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4141 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4201 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4261 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4321 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4381 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4441 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4501 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4561 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4621 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4681 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4741 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4801 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4861 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4921 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4981 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5041 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5101 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5161 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5221 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5281 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5341 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5401 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5461 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5521 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5581 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5641 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5701 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5761 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5821 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5881 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5941 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6001 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6061 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6241 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6361 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7201 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7321 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7501 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7561 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7741 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8041 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8401 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
