LOCUS       V019178                 7079 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019178
VERSION     V019178
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             683..1759
                     /label="SCD(NM_005063)"
                     /note="SCD(NM_005063)"
                     /gene="SCD"
     CDS             1766..1831
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1855..2443
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2509..2557
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2759..3187
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3201..3530
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3597..4391
                     /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    4567..4688
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4737..4753)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4761..4777
                     /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(4785..4815)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4830..4851
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5139..5727)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5898..6758)
                     /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(6759..6863)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgccggc ccacttgctg caggacgata tctctagctc
      721 ctataccacc accaccacca ttacagcgcc tccctccagg gtcctgcaga atggaggaga
      781 taagttggag acgatgcccc tctacttgga agacgacatt cgccctgata taaaagatga
      841 tatatatgac cccacctaca aggataagga aggcccaagc cccaaggttg aatatgtctg
      901 gagaaacatc atccttatgt ctctgctaca cttgggagcc ctgtatggga tcactttgat
      961 tcctacctgc aagttctaca cctggctttg gggggtattc tactattttg tcagtgccct
     1021 gggcataaca gcaggagctc atcgtctgtg gagccaccgc tcttacaaag ctcggctgcc
     1081 cctacggctc tttctgatca ttgccaacac aatggcattc cagaatgatg tctatgaatg
     1141 ggctcgtgac caccgtgccc accacaagtt ttcagaaaca catgctgatc ctcataattc
     1201 ccgacgtggc tttttcttct ctcacgtggg ttggctgctt gtgcgcaaac acccagctgt
     1261 caaagagaag gggagtacgc tagacttgtc tgacctagaa gctgagaaac tggtgatgtt
     1321 ccagaggagg tactacaaac ctggcttgct gatgatgtgc ttcatcctgc ccacgcttgt
     1381 gccctggtat ttctggggtg aaacttttca aaacagtgtg ttcgttgcca ctttcttgcg
     1441 atatgctgtg gtgcttaatg ccacctggct ggtgaacagt gctgcccacc tcttcggata
     1501 tcgtccttat gacaagaaca ttagcccccg ggagaatatc ctggtttcac ttggagctgt
     1561 gggtgagggc ttccacaact accaccactc ctttccctat gactactctg ccagtgagta
     1621 ccgctggcac atcaacttca ccacattctt cattgattgc atggccgccc tcggtctggc
     1681 ctatgaccgg aagaaagtct ccaaggccgc catcttggcc aggattaaaa gaaccggaga
     1741 tggaaactac aagagtggcc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1801 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     1861 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1921 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1981 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2041 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2101 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2161 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2221 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2281 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2341 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2401 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2461 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2521 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2581 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2641 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2701 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2761 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2821 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2881 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2941 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3001 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3061 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3121 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3181 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3361 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3421 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3481 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3541 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3601 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3661 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3721 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3781 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3841 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3901 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3961 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4021 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4081 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4141 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4201 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4261 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4321 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4381 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4441 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4501 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4561 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4681 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6901 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     6961 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7021 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
