LOCUS       V019114                 8189 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019114
VERSION     V019114
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..2869
                     /label="SLC15A2(NM_021082)"
                     /note="SLC15A2(NM_021082)"
                     /gene="SLC15A2"
     CDS             2876..2941
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2965..3553
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3619..3667
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3869..4297
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4311..4640
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4707..5501
                     /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    5677..5798
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5847..5863)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5871..5887
                     /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(5895..5925)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5940..5961
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6249..6837)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7008..7868)
                     /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(7869..7973)
                     /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 ccatgaatcc tttccagaaa aatgagtcca aggaaactct
      721 tttttcacct gtctccattg aagaggtacc acctcgacca cctagccctc caaagaagcc
      781 atctccgaca atctgtggct ccaactatcc actgagcatt gccttcattg tggtgaatga
      841 attctgcgag cgcttttcct attatggaat gaaagctgtg ctgatcctgt atttcctgta
      901 tttcctgcac tggaatgaag atacctccac atctatatac catgccttca gcagcctctg
      961 ttattttact cccatcctgg gagcagccat tgctgactcg tggttgggaa aattcaagac
     1021 aatcatctat ctctccttgg tgtatgtgct tggccatgtg atcaagtcct tgggtgcctt
     1081 accaatactg ggaggacaag tggtacacac agtcctatca ttgatcggcc tgagtctaat
     1141 agctttgggg acaggaggca tcaaaccctg tgtggcagct tttggtggag accagtttga
     1201 agaaaaacat gcagaggaac ggactagata cttctcagtc ttctacctgt ccatcaatgc
     1261 agggagcttg atttctacat ttatcacacc catgctgaga ggagatgtgc aatgttttgg
     1321 agaagactgc tatgcattgg cttttggagt tccaggactg ctcatggtaa ttgcacttgt
     1381 tgtgtttgca atgggaagca aaatatacaa taaaccaccc cctgaaggaa acatagtggc
     1441 tcaagttttc aaatgtatct ggtttgctat ttccaatcgt ttcaagaacc gttctggaga
     1501 cattccaaag cgacagcact ggctagactg ggcggctgag aaatatccaa agcagctcat
     1561 tatggatgta aaggcactga ccagggtact attcctttat atcccattgc ccatgttctg
     1621 ggctcttttg gatcagcagg gttcacgatg gactttgcaa gccatcagga tgaataggaa
     1681 tttggggttt tttgtgcttc agccggacca gatgcaggtt ctaaatcccc ttctggttct
     1741 tatcttcatc ccgttgtttg actttgtcat ttatcgtctg gtctccaagt gtggaattaa
     1801 cttctcatca cttaggaaaa tggctgttgg tatgatccta gcatgcctgg catttgcagt
     1861 tgcggcagct gtagagataa aaataaatga aatggcccca gcccagccag gtccccagga
     1921 ggttttccta caagtcttga atctggcaga tgatgaggtg aaggtgacag tggtgggaaa
     1981 tgaaaacaat tctctgttga tagagtccat caaatccttt cagaaaacac cacactattc
     2041 caaactgcac ctgaaaacaa aaagccagga ttttcacttc cacctgaaat atcacaattt
     2101 gtctctctac actgagcatt ctgtgcagga gaagaactgg tacagtcttg tcattcgtga
     2161 agatgggaac agtatctcca gcatgatggt aaaggataca gaaagcagaa caaccaatgg
     2221 gatgacaacc gtgaggtttg ttaacacttt gcataaagat gtcaacatct ccctgagtac
     2281 agatacctct ctcaatgttg gtgaagacta tggtgtgtct gcttatagaa ctgtgcaaag
     2341 aggagaatac cctgcagtgc actgtagaac agaagataag aacttttctc tgaatttggg
     2401 tcttctagac tttggtgcag catatctgtt tgttattact aataacacca atcagggtct
     2461 tcaggcctgg aagattgaag acattccagc caacaaaatg tccattgcgt ggcagctacc
     2521 acaatatgcc ctggttacag ctggggaggt catgttctct gtcacaggtc ttgagttttc
     2581 ttattctcag gctccctcta gcatgaaatc tgtgctccag gcagcttggc tattgacaat
     2641 tgcagttggg aatatcatcg tgcttgttgt ggcacagttc agtggcctgg tacagtgggc
     2701 cgaattcatt ttgttttcct gcctcctgct ggtgatctgc ctgatcttct ccatcatggg
     2761 ctactactat gttcctgtaa agacagagga tatgcggggt ccagcagata agcacattcc
     2821 tcacatccag gggaacatga tcaaactaga gaccaagaag acaaaactcc tcgaggacta
     2881 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2941 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     3001 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3061 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3121 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3181 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3241 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3301 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     3361 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3421 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3481 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3541 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     3601 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3661 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3721 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3781 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     3841 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3901 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3961 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4021 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4081 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4141 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4201 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     4261 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4321 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4441 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4501 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4561 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4621 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4681 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4741 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4801 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4861 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4921 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4981 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5041 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5101 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5161 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5221 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5281 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5341 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5401 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5461 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     5521 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5581 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5641 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5701 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5761 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5821 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5881 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5941 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6001 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6061 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6121 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6181 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6241 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6301 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6361 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6421 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6481 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6541 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6601 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6661 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6961 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7141 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7321 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7621 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7861 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7981 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     8041 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     8101 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     8161 ccgacaattg catgaagaat ctgcttagg
//
