LOCUS       V032100                 6729 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032100
VERSION     V032100
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..1411
                     /label="SENP8(NM_145204)"
                     /note="SENP8(NM_145204)"
                     /gene="SENP8"
     CDS             1412..1435
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     misc_feature    1505..2093
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2159..2207
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2409..2837
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2851..3180
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3247..4041
                     /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    4217..4338
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4387..4403)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4411..4427
                     /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(4435..4465)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4480..4501
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4789..5377)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5548..6408)
                     /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(6409..6513)
                     /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 tcggatccag ccaccatgga
      781 ccccgtagtc ttgagttaca tggacagtct actgcggcaa tcagatgtct cactattgga
      841 tccgccaagc tggctcaatg accatattat tgggtttgcg tttgagtact ttgccaacag
      901 tcagtttcat gactgctctg atcacgtcag tttcatcagc cctgaagtca cccagttcat
      961 caagtgcact agcaacccag cagagattgc catgttcctt gaaccactgg acctccccaa
     1021 caagagagtt gtatttttag ccatcaatga taactccaac caggcagctg gaggaaccca
     1081 ctggagttta ttggtctacc tccaagataa aaatagcttt tttcattatg attcccatag
     1141 caggagcaac tcagttcacg caaagcaggt agcagagaaa ctggaggctt tcttaggcag
     1201 aaaaggagac aaactggcct ttgtggaaga gaaagcccct gcccaacaaa acagctatga
     1261 ctgtgggatg tacgtgatat gtaacactga ggccttgtgt cagaacttct ttaggcaaca
     1321 gacagaatca ctgctgcagc tactcacccc tgcatacatc acaaagaaga ggggagaatg
     1381 gaaagatctc attaccacac ttgctaaaaa gtggagccac ccgcagttcg aaaagtagct
     1441 cgagtctaga gggcccctta agggttcgat ccctaccggt tagtaatgag tttgatatct
     1501 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1561 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1621 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     1681 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     1741 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     1801 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1861 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     1921 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     1981 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2041 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2101 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2161 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2221 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     2281 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     2341 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     2401 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     2581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2761 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     2821 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2881 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2941 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3181 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3241 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3301 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3361 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3421 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3481 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3541 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3601 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3661 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3721 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3781 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3841 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3901 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3961 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4021 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4081 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4141 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4201 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4321 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4381 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4441 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4501 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4981 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5221 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5341 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5401 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5461 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5521 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5581 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5641 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5701 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5761 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5821 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5881 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5941 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6001 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6061 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6121 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6181 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6241 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6301 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6361 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6421 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6481 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6541 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     6601 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     6661 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     6721 ctgcttagg
//
