LOCUS       V037339                 9638 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037339
VERSION     V037339
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"
     CDS             775..3498
                     /label="PSMD2(NM_002808)"
                     /note="PSMD2(NM_002808)"
                     /gene="PSMD2"
     CDS             3499..3564
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3580..3636
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3637..4347
                     /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"
     misc_feature    4414..5002
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5068..5116
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5318..5746
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5760..6089
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6156..6950
                     /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    7126..7247
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7296..7312)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7320..7336
                     /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(7344..7374)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7389..7410
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7698..8286)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8457..9317)
                     /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(9318..9422)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggag
      781 gagggaggcc gggacaaggc gccggtgcag ccccagcagt ctccagcggc ggcccccggc
      841 ggcacggacg agaagccgag cggcaaggag cggcgggatg ccggggacaa ggacaaagaa
      901 caggagctgt ctgaagagga taaacagctt caagatgaac tggagatgct cgtggaacga
      961 ctaggggaga aggatacatc cctgtatcga ccagcgctgg aggaattgcg aaggcagatt
     1021 cgttcttcta caacttccat gacttcagtg cccaagcctc tcaaatttct gcgtccacac
     1081 tatggcaaac tgaaggaaat ctatgagaac atggcccctg gggagaataa gcgttttgct
     1141 gctgacatca tctccgtttt ggccatgacc atgagtgggg agcgtgagtg cctcaagtat
     1201 cggctagtgg gctcccagga ggaattggca tcatggggtc atgagtatgt caggcatctg
     1261 gcaggagaag tggctaagga gtggcaggag ctggatgacg cagagaaggt ccagcgggag
     1321 cctctgctca ctctggtgaa ggaaatcgtc ccctataaca tggcccacaa tgcagagcat
     1381 gaggcttgcg acctgcttat ggaaattgag caggtggaca tgctggagaa ggacattgat
     1441 gaaaatgcat atgcaaaggt ctgcctttat ctcaccagtt gtgtgaatta cgtgcctgag
     1501 cctgagaact cagccctact gcgttgtgcc ctgggtgtgt tccgaaagtt tagccgcttc
     1561 cctgaagctc tgagattggc attgatgctc aatgacatgg agttggtaga agacatcttc
     1621 acctcctgca aggatgtggt agtacagaaa cagatggcat tcatgctagg ccggcatggg
     1681 gtgttcctgg agctgagtga agatgtcgag gagtatgagg acctgacaga gatcatgtcc
     1741 aatgtacagc tcaacagcaa cttcttggcc ttagctcggg agctggacat catggagccc
     1801 aaggtgcctg atgacatcta caaaacccac ctagagaaca acaggtttgg gggcagtggc
     1861 tctcaggtgg actctgcccg catgaacctg gcctcctctt ttgtgaatgg ctttgtgaat
     1921 gcagcttttg gccaagacaa gctgctaaca gatgatggca acaaatggct ttacaagaac
     1981 aaggaccacg gaatgttgag tgcagctgca tctcttggga tgattctgct gtgggatgtg
     2041 gatggtggcc tcacccagat tgacaagtac ctgtactcct ctgaggacta cattaagtca
     2101 ggagctcttc ttgcctgtgg catagtgaac tctggggtcc ggaatgagtg tgaccctgct
     2161 ctggcactgc tctcagacta tgttctccac aacagcaaca ccatgagact tggttccatc
     2221 tttgggctag gcttggctta tgctggctca aatcgtgaag atgtcctaac actgctgctg
     2281 cctgtgatgg gagattcaaa gtccagcatg gaggtggcag gtgtcacagc tttagcctgt
     2341 ggaatgatag cagtagggtc ctgcaatgga gatgtaactt ccactatcct tcagaccatc
     2401 atggagaagt cagagactga gctcaaggat acttatgctc gttggcttcc tcttggactg
     2461 ggtctcaacc acctggggaa gggtgaggcc atcgaggcaa tcctggctgc actggaggtt
     2521 gtgtcagagc cattccgcag ttttgccaac acactggtgg atgtgtgtgc atatgcaggc
     2581 tctgggaatg tgctgaaggt gcagcagctg ctccacattt gtagcgaaca ctttgactcc
     2641 aaagagaagg aggaagacaa agacaagaag gaaaagaaag acaaggacaa gaaggaagcc
     2701 cctgctgaca tgggagcaca tcagggagtg gctgttctgg ggattgccct tattgctatg
     2761 ggggaggaga ttggtgcaga gatggcatta cgaacctttg gccacttgct gagatatggg
     2821 gagcctacac tccggagggc tgtaccttta gcactggccc tcatctctgt ttcaaatcca
     2881 cgactcaaca tcctggatac cctaagcaaa ttctctcatg atgctgatcc agaagtttcc
     2941 tataactcca tttttgccat gggcatggtg ggcagtggta ccaataatgc ccgtctggct
     3001 gcaatgctgc gccagttagc tcaatatcat gccaaggacc caaacaacct cttcatggtg
     3061 cgcttggcac agggcctgac acatttaggg aagggcaccc ttaccctctg cccctaccac
     3121 agcgaccggc agcttatgag ccaggtggcc gtggctggac tgctcactgt gcttgtctct
     3181 ttcctggatg ttcgaaacat tattctaggc aaatcacact atgtattgta tgggctggtg
     3241 gctgccatgc agccccgaat gctggttacg tttgatgagg agctgcggcc attgccagtg
     3301 tctgtccgtg tgggccaggc agtggatgtg gtgggccagg ctggcaagcc gaagactatc
     3361 acagggttcc agacgcatac aaccccagtg ttgttggccc acggggaacg ggcagaattg
     3421 gccactgagg agtttcttcc tgttaccccc attctggaag gttttgttat ccttcggaag
     3481 aaccccaatt atgatctcga ctacaaagac catgacggtg attataaaga tcatgacatc
     3541 gactacaagg atgacgatga caaggaattc ggaagcggag ctactaactt cagcctgctg
     3601 aagcaggctg gcgacgtgga ggagaaccct ggacctatgg tgagcaaggg cgaggaggat
     3661 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3721 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3781 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3841 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3901 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3961 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     4021 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     4081 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     4141 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     4201 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     4261 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     4321 ggcggcatgg acgagctgta caagtaactc gagtctagag ggccccttaa gggttcgatc
     4381 cctaccggtt agtaatgagt ttgatatctc gacaatcaac ctctggatta caaaatttgt
     4441 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4501 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4561 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4621 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4681 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4741 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4801 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4861 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4921 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4981 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     5041 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     5101 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5161 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5221 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5281 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     5341 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5401 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5461 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5521 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5581 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5641 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     5701 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6001 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     6061 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     6121 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6181 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6241 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6301 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     6361 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6421 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6481 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6541 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6601 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6661 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6721 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6781 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6841 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6901 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6961 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     7021 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     7081 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     7141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     7261 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7861 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8101 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     8161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9421 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     9481 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     9541 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     9601 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
