LOCUS       V033047                 9197 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033047
VERSION     V033047
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..3123
                     /label="TRIM67(NM_001004342)"
                     /note="TRIM67(NM_001004342)"
                     /gene="TRIM67"
     CDS             3139..3195
                     /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             3196..3906
                     /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    3973..4561
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4627..4675
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4877..5305
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5319..5648
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5715..6509
                     /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    6685..6806
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6855..6871)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6879..6895
                     /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(6903..6933)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6948..6969
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7257..7845)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8016..8876)
                     /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(8877..8981)
                     /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 gaagagctga agtgtcccgt gtgcggctct ctgtttcggg agcctatcat cctgccctgt
      841 tcccacaatg tctgcctgcc ttgcgctcgc accatcgcgg tgcagacccc ggacggtgag
      901 cagcacctgc cccagccgct cctgctttcc cggggatcgg ggctgcaggc gggcgccgcc
      961 gccgctgcct ctctggagca cgacgctgcg gctggcccgg cctgcggcgg tgcaggcggg
     1021 agtgcagctg gcggcctcgg cggcggtgcg ggaggtggcg gagaccacgc ggacaagctc
     1081 agcttgtaca gcgagacaga cagcggctac gggtcctaca ccccgagcct caagtccccc
     1141 aacggggttc gcgtgctgcc catggtgccc gcaccacccg gctcctcggc tgcggcggct
     1201 cggggtgccg cctgctcctc gctgtcctcg tcttcgagct ccatcacgtg cccgcagtgc
     1261 caccgcagcg catccctgga ccaccgcggc ctgcgcggct tccagcgcaa ccggctgctc
     1321 gaggccatcg tgcagcggta ccagcagggc cgcggggccg tgccggggac gtctgcagcc
     1381 gcggcggtgg ccatctgcca gctgtgcgac cgcaccccgc cagagccagc agccacgctc
     1441 tgcgagcagt gcgacgtcct ctactgctct gcctgccagc tcaagtgcca tccatcccgg
     1501 ggacccttcg ccaagcatcg cctggtgcag ccgccgccgc cgccgccgcc gcccgccgag
     1561 gcagcctccg ggcccactgg caccgcccag ggcgccccca gcggaggcgg cggctgcaag
     1621 agcccgggag gcgcgggggc gggggcgact gggggcagca cggcccgcaa gttccccacg
     1681 tgtcccgagc atgaaatgga gaactacagc atgtactgcg tgagctgtcg aaccccggtg
     1741 tgttatctgt gcctggagga gggccggcac gccaagcacg aggtgaagcc gctgggggcc
     1801 atgtggaagc agcacaaggc acaactatct caggccttaa atggagtttc agataaggca
     1861 aaggaagcaa aggagtttct ggttcagcta aagaacatat tgcagcagat ccaggaaaac
     1921 ggactggact acgaagcctg cctcgttgct cagtgtgatg cccttgtgga tgctttaact
     1981 cgtcagaaag ccaagctgct caccaaggtg actaaagaga gggaacacaa gttgaagatg
     2041 gtttgggacc agatcaatca ctgcacattg aagctgcgtc agtccaccgg actgatggag
     2101 tactgcctgg aggtgatcaa ggagaacgac ccctccgggt tcttacagat ctcagatgct
     2161 ctgatcaagc gcgtccaggt gtctcaggag cagtgggtca aaggcgccct ggagccgaaa
     2221 gtgtctgcgg agtttgatct gactttggac agcgagccgc tgctgcaggc catccaccag
     2281 ctggacttca ttcagatgaa atgtagggtg ccacccgtcc ccctactgca gctggagaaa
     2341 tgctgcaccc gtaacaacag cgtcacgctg gcctggagga tgccaccctt cacccacagc
     2401 cccgtggacg gctacatcct ggagctggac gacggtgccg ggggacagtt ccgggaagtg
     2461 tacgtcggta aggagacttt gtgtaccatc gacggtcttc acttcaacag cacctacaac
     2521 gcccgagtca aagctttcaa ctcttctggt gtcgggcctt acagtaaaac tgtcgtcctg
     2581 cagacatccg atgtggcctg gttcacattt gaccccaact ctgggcatcg ggacatcatt
     2641 ttatccaatg acaaccagac agccacctgc agcagctatg acgaccgggt ggtgctgggc
     2701 acagctgcgt tctccaaggg cgtgcactac tgggagctgc acgtggaccg gtacgacaac
     2761 cacccagacc ccgccttcgg ggtggccagg gccagcgtgg tcaaggacat gatgctgggc
     2821 aaggatgaca aggcctgggc catgtatgtg gacaacaacc gcagctggtt catgcactgc
     2881 aactcccaca ccaacaggac ggaaggtggc gtgtgcaagg gggccaccgt gggcgtgctg
     2941 ctggacctga ataagcacac tctcaccttc ttcatcaacg ggcagcagca gggccccaca
     3001 gccttcagcc acgtggacgg ggtcttcatg ccagccctca gcctcaaccg caacgtgcag
     3061 gtcaccctgc acacaggatt ggaagtgccg actaacctgg ggcggccaaa gctgtcaggc
     3121 aatgaattcg gaagcggagc tactaacttc agcctgctga agcaggctgg cgacgtggag
     3181 gagaaccctg gacctatggt gagcaagggc gaggaggata acatggccat catcaaggag
     3241 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3301 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3361 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     3421 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     3481 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     3541 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3601 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     3661 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     3721 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     3781 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     3841 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     3901 aagtaactcg agtctagagg gccccttaag ggttcgatcc ctaccggtta gtaatgagtt
     3961 tgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4021 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4081 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4141 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4201 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4261 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4321 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4381 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4441 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4501 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4561 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4621 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     4681 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4741 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4801 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     4861 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4921 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4981 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     5041 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5101 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5161 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5221 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     5281 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5581 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5641 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5701 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5761 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5821 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5881 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5941 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6001 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6061 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6121 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6181 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6241 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6301 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6361 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6421 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6481 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6541 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6601 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6661 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7081 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7441 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7681 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7921 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7981 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8041 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8101 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8161 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8221 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8281 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8341 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8401 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8461 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8521 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8581 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8641 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8701 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8761 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8821 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8941 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9001 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     9061 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     9121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     9181 tgaagaatct gcttagg
//
