LOCUS       V030829                 8031 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030829
VERSION     V030829
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /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             1500..2711
                     /label="Fcgr1(NM_010186)"
                     /note="Fcgr1(NM_010186)"
                     /gene="Fcgr1"
     CDS             2718..2783
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2807..3395
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3461..3509
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3711..4139
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4153..4482
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4549..5343
                     /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    5519..5640
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5689..5705)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5713..5729
                     /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(5737..5767)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5782..5803
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6091..6679)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6850..7710)
                     /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(7711..7815)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tgattcttac cagctttgga gatgacatgt ggcttctaac aactctgcta ctttgggttc
     1561 cagtcggtgg ggaagtggtt aatgccacca aggctgtgat caccttgcag cctccatggg
     1621 tcagtatttt ccagaaggaa aatgtcactt tatggtgtga ggggcctcac ctgcctggag
     1681 acagttccac acaatggttt atcaacggaa cagccgttca gatctccacg cctagttata
     1741 gcatcccaga ggccagtttt caggacagtg gcgaatacag gtgtcagata ggttcctcaa
     1801 tgccaagtga ccctgtgcag ttgcaaatcc acaatgattg gctgctactc caggcctccc
     1861 gcagagtcct cacagaagga gaacccctgg ccttgaggtg tcacggatgg aagaataaac
     1921 tggtgtacaa tgtggttttc tatagaaatg gaaaatcctt tcagttttct tcagattcgg
     1981 aggtcgccat tctgaaaacc aacctgagtc acagcggcat ctaccactgc tcaggcacgg
     2041 gaagacaccg ctacacatct gcaggagtgt ccatcacggt gaaagagctg tttaccacgc
     2101 cagtgctgag agcatccgtg tcatctccct tcccggaggg gagtctggtc accctgaact
     2161 gtgagacgaa tttgctcctg cagagacccg gcttacagct tcacttctcc ttctacgtgg
     2221 gcagcaagat cctggagtac aggaacacat cctcagagta ccatatagca agggcggaaa
     2281 gagaagatgc tggattctac tggtgtgagg tagccacgga ggacagcagt gtccttaagc
     2341 gcagccctga gttggagctc caagtgcttg gtccccagtc atcagctcct gtctggtttc
     2401 acatcctgtt ttatctgtca gtgggaataa tgttttcgtt gaacacggtt ctctatgtga
     2461 aaatacacag gctgcagaga gagaagaaat acaacttaga agtccctttg gtttctgagc
     2521 agggaaagaa agcaaattcc tttcagcaag ttagaagcga tggcgtgtat gaagaagtaa
     2581 cagccactgc gagccagacc acaccaaaag aagcgcccga tggacctcga agctcagtgg
     2641 gtgactgtgg acccgagcag cctgaacccc ttcctcccag tgacagtact ggggcacaaa
     2701 cttcccaaag tctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg
     2761 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2821 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2881 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2941 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3001 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3061 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3121 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3181 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3241 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3301 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3361 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3421 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3481 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3541 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3601 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3661 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3721 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3781 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3841 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3901 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3961 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4021 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4081 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4141 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4201 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4321 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4381 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4441 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4501 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4561 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4621 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4681 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4741 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4801 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4861 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4921 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4981 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5041 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5101 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5161 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5221 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5281 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5341 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5401 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5461 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5521 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5581 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5641 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5701 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5761 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5821 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5881 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5941 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6001 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7861 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7921 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7981 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
