LOCUS       V044704                 8028 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044704
VERSION     V044704
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..2708
                     /label="IDO1(NM_002164)"
                     /note="IDO1(NM_002164)"
                     /gene="IDO1"
     CDS             2715..2780
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2804..3392
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3458..3506
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3708..4136
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4150..4479
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4546..5340
                     /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    5516..5637
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5686..5702)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5710..5726
                     /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(5734..5764)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5779..5800
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6088..6676)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6847..7707)
                     /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(7708..7812)
                     /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 tggcacacgc tatggaaaac tcctggacaa tcagtaaaga gtaccatatt gatgaagaag
     1561 tgggctttgc tctgccaaat ccacaggaaa atctacctga tttttataat gactggatgt
     1621 tcattgctaa acatctgcct gatctcatag agtctggcca gcttcgagaa agagttgaga
     1681 agttaaacat gctcagcatt gatcatctca cagaccacaa gtcacagcgc cttgcacgtc
     1741 tagttctggg atgcatcacc atggcatatg tgtggggcaa aggtcatgga gatgtccgta
     1801 aggtcttgcc aagaaatatt gctgttcctt actgccaact ctccaagaaa ctggaactgc
     1861 ctcctatttt ggtttatgca gactgtgtct tggcaaactg gaagaaaaag gatcctaata
     1921 agcccctgac ttatgagaac atggacgttt tgttctcatt tcgtgatgga gactgcagta
     1981 aaggattctt cctggtctct ctattggtgg aaatagcagc tgcttctgca atcaaagtaa
     2041 ttcctactgt attcaaggca atgcaaatgc aagaacggga cactttgcta aaggcgctgt
     2101 tggaaatagc ttcttgcttg gagaaagccc ttcaagtgtt tcaccaaatc cacgatcatg
     2161 tgaacccaaa agcatttttc agtgttcttc gcatatattt gtctggctgg aaaggcaacc
     2221 cccagctatc agacggtctg gtgtatgaag ggttctggga agacccaaag gagtttgcag
     2281 ggggcagtgc aggccaaagc agcgtctttc agtgctttga cgtcctgctg ggcatccagc
     2341 agactgctgg tggaggacat gctgctcagt tcctccagga catgagaaga tatatgccac
     2401 cagctcacag gaacttcctg tgctcattag agtcaaatcc ctcagtccgt gagtttgtcc
     2461 tttcaaaagg tgatgctggc ctgcgggaag cttatgacgc ctgtgtgaaa gctctggtct
     2521 ccctgaggag ctaccatctg caaatcgtga ctaagtacat cctgattcct gcaagccagc
     2581 agccaaagga gaataagacc tctgaagacc cttcaaaact ggaagccaaa ggaactggag
     2641 gcactgattt aatgaatttc ctgaagactg taagaagtac aactgagaaa tcccttttga
     2701 aggaaggtct cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2761 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2821 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2881 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2941 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3001 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3061 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3121 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3181 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3241 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3301 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3361 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3421 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3481 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3541 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3601 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3661 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3721 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3781 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3841 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3901 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3961 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4021 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     4081 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4141 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4201 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4261 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4321 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4381 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4441 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4501 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4561 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4621 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4681 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4741 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4801 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4861 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4921 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4981 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5041 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5101 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5161 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5221 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5281 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5341 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5401 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5461 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5521 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5581 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5641 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5701 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5761 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5821 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5881 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7801 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7861 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7921 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7981 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
