LOCUS       V046315                 9894 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046315
VERSION     V046315
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             776..3820
                     /label="PTPRN2(NM_002847)"
                     /note="PTPRN2(NM_002847)"
                     /gene="PTPRN2"
     CDS             3836..3892
                     /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             3893..4603
                     /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    4670..5258
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5324..5372
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5574..6002
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6016..6345
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6412..7206
                     /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    7382..7503
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7552..7568)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7576..7592
                     /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(7600..7630)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7645..7666
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7954..8542)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8713..9573)
                     /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(9574..9678)
                     /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 tcggatcccg ccaccatggg
      781 gccgccgctc ccgctgctgc tgctgctact gctgctgctg ccgccacgcg tcctgcctgc
      841 cgccccttcg tccgtccccc gcggccggca gctcccgggg cgtctgggct gcctgctcga
      901 ggagggcctc tgcggagcgt ccgaggcctg tgtgaacgat ggagtgtttg gaaggtgcca
      961 gaaggttccg gcaatggact tttaccgcta cgaggtgtcg cccgtggccc tgcagcgcct
     1021 gcgcgtggcg ttgcagaagc tttccggcac aggtttcacg tggcaggatg actatactca
     1081 gtatgtgatg gaccaggaac ttgcagacct cccgaaaacc tacctgaggc gtcctgaagc
     1141 atccagccca gccaggccct caaaacacag cgttggcagc gagaggaggt acagtcggga
     1201 gggcggtgct gccctggcca acgccctccg acgccacctg cccttcctgg aggccctgtc
     1261 ccaggcccca gcctcagacg tgctcgccag gacccatacg gcgcaggaca gaccccccgc
     1321 tgagggtgat gaccgcttct ccgagagcat cctgacctat gtggcccaca cgtctgcgct
     1381 gacctaccct cccgggtccc ggacccagct ccgcgaggac ctcctgccgc ggaccctcgg
     1441 ccagctccag ccagatgagc tcagccctaa ggtggacagt ggtgtggaca gacaccatct
     1501 gatggcggcc ctcagtgcct atgctgccca gaggccccca gctccccccg gggagggcag
     1561 cctggagcca cagtaccttc tgcgtgcacc ctcaagaatg cccaggcctt tgctggcacc
     1621 agccgccccc cagaagtggc cttcacctct gggagattcc gaagacccct ccagcacagg
     1681 cgatggagca cggattcata ccctcctgaa ggacctgcag aggcagccgg ctgaggtgag
     1741 gggcctgagt ggcctggagc tggacggcat ggctgagctg atggctggcc tgatgcaagg
     1801 cgtggaccat ggagtagctc gaggcagccc tgggagagcg gccctgggag agtctggaga
     1861 acaggcggat ggccccaagg ccaccctccg tggagacagc tttccagatg acggagtgca
     1921 ggacgacgat gatagacttt accaagaggt ccatcgtctg agtgccacac tcgggggcct
     1981 cctgcaggac cacgggtctc gactcttacc tggagccctc ccctttgcaa ggcccctcga
     2041 catggagagg aagaagtccg agcaccctga gtcttccctg tcttcagaag aggagactgc
     2101 cggagtggag aacgtcaaga gccagacgta ttccaaagat ctgctggggc agcagccgca
     2161 ttcggagccc ggggccgctg cgtttgggga gctccaaaac cagatgcctg ggccctcgaa
     2221 ggaggagcag agccttccag cgggtgctca ggaggccctc agcgacggcc tgcaattgga
     2281 ggtccagcct tccgaggaag aggcgcgggg ctacatcgtg acagacagag accccctgcg
     2341 ccccgaggaa ggaaggcggc tggtggagga cgtcgcccgc ctcctgcagg tgcccagcag
     2401 tgcgttcgct gacgtggagg ttctcggacc agcagtgacc ttcaaagtga gcgccaatgt
     2461 ccaaaacgtg accactgagg atgtggagaa ggccacagtt gacaacaaag acaaactgga
     2521 ggaaacctct ggactgaaaa ttcttcaaac cggagtcggg tcgaaaagca aactcaagtt
     2581 cctgcctcct caggcggagc aagaagactc caccaagttc atcgcgctca ccctggtctc
     2641 cctcgcctgc atcctgggcg tcctcctggc ctctggcctc atctactgcc tccgccatag
     2701 ctctcagcac aggctgaagg agaagctctc gggactaggg ggcgacccag gtgcagatgc
     2761 cactgccgcc taccaggagc tgtgccgcca gcgtatggcc acgcggccac cagaccgacc
     2821 tgagggcccg cacacgtcac gcatcagcag cgtctcatcc cagttcagcg acgggccgat
     2881 ccccagcccc tccgcacgca gcagcgcctc atcctggtcc gaggagcctg tgcagtccaa
     2941 catggacatc tccaccggcc acatgatcct gtcctacatg gaggaccacc tgaagaacaa
     3001 gaaccggctg gagaaggagt gggaagcgct gtgcgcctac caggcggagc ccaacagctc
     3061 gttcgtggcc cagagggagg agaacgtgcc caagaaccgc tccctggccg tgctgaccta
     3121 tgaccactcc cgggtcctgc tgaaggcgga gaacagccac agccactcag actacatcaa
     3181 cgctagcccc atcatggatc acgacccgag gaaccccgcg tacatcgcca cccagggacc
     3241 gctgcccgcc accgtggctg acttttggca gatggtgtgg gagagcggct gcgtggtgat
     3301 cgtcatgctg acacccctcg cggagaacgg cgtccggcag tgctaccact actggccgga
     3361 tgaaggctcc aatctctacc acatctatga ggtgaacctg gtctccgagc acatctggtg
     3421 tgaggacttc ctggtgagga gcttctatct gaagaacctg cagaccaacg agacgcgcac
     3481 cgtgacgcag ttccacttcc tgagttggta tgaccgagga gtcccttcct cctcaaggtc
     3541 cctcctggac ttccgcagaa aagtaaacaa gtgctacagg ggccgttctt gtccaataat
     3601 tgttcattgc agtgacggtg caggccggag cggcacctac gtcctgatcg acatggttct
     3661 caacaagatg gccaaaggtg ctaaagagat tgatatcgca gcgaccctgg agcacttgag
     3721 ggaccagaga cccggcatgg tccagacgaa ggagcagttt gagttcgcgc tgacagccgt
     3781 ggctgaggag gtgaacgcca tcctcaaggc ccttccccag gaattcggaa gcggagctac
     3841 taacttcagc ctgctgaagc aggctggcga cgtggaggag aaccctggac ctatggtgag
     3901 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     3961 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4021 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4081 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4141 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4201 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     4261 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     4321 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     4381 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     4441 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     4501 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     4561 gggccgccac tccaccggcg gcatggacga gctgtacaag taactcgagt ctagagggcc
     4621 ccttaagggt tcgatcccta ccggttagta atgagtttga tatctcgaca atcaacctct
     4681 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4741 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4801 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4861 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4921 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4981 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5041 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     5101 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5161 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5221 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     5281 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5341 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5401 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5461 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5521 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     5581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5701 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     5761 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5881 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5941 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6001 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     6061 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     6121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6181 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     6301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     6361 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     6421 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6481 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6541 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     6601 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6661 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6721 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6781 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6841 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6901 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     6961 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7021 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7081 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7141 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7201 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     7261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7381 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7501 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7561 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7621 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7741 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     8161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8881 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     9001 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     9061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     9121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9241 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9301 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9421 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9541 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9661 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9721 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     9781 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     9841 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
