LOCUS       V037307                 8789 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037307
VERSION     V037307
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..2715
                     /label="EEF1D(NM_001130053)"
                     /note="EEF1D(NM_001130053)"
                     /gene="EEF1D"
     CDS             2731..2787
                     /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             2788..3498
                     /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    3565..4153
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4219..4267
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4469..4897
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4911..5240
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5307..6101
                     /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    6277..6398
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6447..6463)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6471..6487
                     /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(6495..6525)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6540..6561
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6849..7437)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7608..8468)
                     /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(8469..8573)
                     /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 caccatgagg
      781 agcgggaagg cctcctgcac cctggagacc gtgtgggaag acaagcacaa gtatgaggag
      841 gccgagcggc gcttctacga acacgaggcc acacaggcgg ccgcctccgc ccagcagctg
      901 ccagccgagg ggccagccat gaatgggccc ggccaggacg accctgagga cgctgatgag
      961 gcggaagccc ctgacggcgg cagcaggcgt gatcccagga agagccagga cagcaggaag
     1021 cccctgcaga aaaagaggaa gcgctccccc aagagcgggc tcggccccgc ggacctggcc
     1081 ctcctgggcc tctcggccga acgcgtgtgg ctggacaagt cacttttcga ccaggcagag
     1141 agctcctacc gccagaagct ggcagatgtg gctgcccagg cagcctggcc tcctgccttg
     1201 gccccttggg gtctctgcac ccatggaaac caggtggcct gccaccacgt gacctggggg
     1261 atctgggtca acaagtcctc cttcgaccag gctgagcggg ccttcgtgga gtggtctcag
     1321 gccctgttgc tggcccccga cggcagccgc aggcagggga ctcccaacac aggccagcag
     1381 gtggccgtcc ccgacctggc ccaccagccc agcccaccgg tcaatggcca gcccccgctg
     1441 ggcagcctgc aggcactggt tcgggaggtg tggctggaga agccccggta tgatgcagcc
     1501 gagaggggct tctacgaggc cctgtttgac ggccatcccc cagggaaggt gcgcctgcaa
     1561 gagcgagccg gcctggccga gggtgcccgg cggggccgca gagaccggcg gggccgcaac
     1621 atcttaggga acaagcgggc cgggctgcga cgggccgatg gggaggcccc ctctgccttg
     1681 ccctactgtt acttcctgca gaaggatgca gaggccccct ggctcagcaa gcctgcctac
     1741 gacagcgccg agtgccgcca ccacgctgcc gaggccctgc gggtggcctg gtgcctcgaa
     1801 gctgcctccc tgtctcaccg acccggtcct cggtctggcc tgtccgtgtc cagcctgaga
     1861 cccaacagaa aaatggctac aaacttccta gcacatgaga agatctggtt cgacaagttc
     1921 aaatatgacg acgcagaaag gagattctac gagcagatga acgggcctgt ggcaggtgcc
     1981 tcccgccagg agaacggcgc cagcgtgatc ctccgtgaca ttgcgagagc cagagagaac
     2041 atccagaaat ccctggctgg aagctcaggc cccggggcct ccagcggcac cagcggagac
     2101 cacggtgagc tcgtcgtccg gattgccagt ctggaagtgg agaaccagag tctgcgtggc
     2161 gtggtacagg agctgcagca ggccatctcc aagctggagg cccggctgaa cgtgctggag
     2221 aagagctcgc ctggccaccg ggccacggcc ccacagaccc agcacgtatc tcccatgcgc
     2281 caagtggagc ccccagccaa gaagccagcc acaccagcag aggatgacga ggatgatgac
     2341 attgacctgt ttggcagtga caatgaggag gaggacaagg aggcggcaca gctgcgggag
     2401 gagcggctac ggcagtacgc ggagaagaag gccaagaagc ctgcactggt ggccaagtcc
     2461 tccatcctgc tggatgtcaa gccttgggat gatgagacgg acatggccca gctggaggcc
     2521 tgtgtgcgct ctatccagct ggacgggctg gtctgggggg cttccaagct ggtgcccgtg
     2581 ggctacggta tccggaagct acagattcag tgtgtggtgg aggacgacaa ggtggggaca
     2641 gacttgctgg aggaggagat caccaagttt gaggagcacg tgcagagtgt cgatatcgca
     2701 gctttcaaca agatcgaatt cggaagcgga gctactaact tcagcctgct gaagcaggct
     2761 ggcgacgtgg aggagaaccc tggacctatg gtgagcaagg gcgaggagga taacatggcc
     2821 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2881 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2941 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3001 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3061 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3121 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     3181 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3241 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3301 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     3361 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3421 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     3481 gacgagctgt acaagtaact cgagtctaga gggcccctta agggttcgat ccctaccggt
     3541 tagtaatgag tttgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     3601 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3661 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3721 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3781 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3841 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3901 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     3961 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4021 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4081 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4141 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     4201 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     4261 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4321 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4381 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     4441 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4921 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     5281 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5341 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5401 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5461 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5521 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5581 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5641 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5701 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5761 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5821 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5881 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5941 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6001 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6061 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     6121 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6181 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6241 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     6301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6361 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6421 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7021 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     7081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7261 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8581 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     8641 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     8701 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     8761 ccgacaattg catgaagaat ctgcttagg
//
