LOCUS       V045103                 7743 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045103
VERSION     V045103
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2423
                     /label="Slc22a20(NM_198650)"
                     /note="Slc22a20(NM_198650)"
                     /gene="Slc22a20"
     CDS             2430..2495
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2519..3107
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3173..3221
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3423..3851
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3865..4194
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4261..5055
                     /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    5231..5352
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5401..5417)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5425..5441
                     /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(5449..5479)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5494..5515
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5803..6391)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6562..7422)
                     /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(7423..7527)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cttcacagac ctgctggatg
      781 ccctgggagg cgtgggtcgc ttccagcttg tctatacagc cttgctgctg ctgccctgtg
      841 ggctgctggc ttgccatacc ttcctacaga acttcacggc tgctgcaccc cctcaccact
      901 gccaacatcc tgccaactac acagaaccta ccaccaatgt ctcaggggtc tggctgaggg
      961 ctgccatacc cctgaaccag catggggacc ccgagccgtg ccggcgctat gtagaacctc
     1021 agtgggccct tctgaaaccc aacgcctcct cccatggggt ggccaccgag ggctgcaagg
     1081 atggttgggt ctacgaccga agcattttcc catctaccat tgtgatggag tgggacctgg
     1141 tgtgtgaggc ccgcaccctt cgtgacctgg ctcagtccat ctacatgtcc ggggtgttgg
     1201 tgggagccgc cttgtttggt ggtcttgctg acaggttggg tcgcaaggcc ccactggtgt
     1261 ggtcgtacct gcagctggca gtttcggggg ctgccacagc gtatgtgggc tccttcagtg
     1321 cctactgcgt cttccgcttc ctgatgggca tgaccttctc cggcatcatc cttaactccc
     1381 tctccctggt tgtagaatgg atgccaacca gaggccgtac agtagcaggc atcttgttgg
     1441 gtttctcctt caccttgggc cagctcatcc tggctggcgt ggcatacctg atccggccct
     1501 ggcggtggct gcagtttgct gtgtctgctc cttttctggt ctttttcctc tattcttggt
     1561 ggcttccaga gtcatcccga tggctcctcc ttcacggcaa ggcccagcaa gctgtgcaga
     1621 accttcagaa ggtggccatg atgaatggca ggaaggcgga aggggagagg ctgactacag
     1681 aggtggtgag ctcctacatc caggatgagt ttgcaagtgt ccgcacctcc aactccatct
     1741 tggacctctt tagaacccct gccatcagaa gggtcacgtg ctgtctcatg ggggtctggt
     1801 tctctaattc tgtggcttac tacggcctgg ccatggacct gcagaagttt gggctcagca
     1861 tctacctggt acaggctctg tttgggatca tcgatatccc ggccatgctg gtcgccacca
     1921 ccacgatgat ttatgtggga cgacgggcca cagtgtcctc cttcctcata ctagctgggc
     1981 tcatggtcat tgccaacatg tttatgcctg aggatctgca gaccttgcgg acagtgcagg
     2041 cagcgctggg caaaggctgc ctggccagct ccttcatctg cgtgtatctg ttcacgggag
     2101 aactctatcc cacagagatc aggcagatgg ggatgggatt tgcttctgtc aatgcccgcc
     2161 tcggaggctt ggtggcaccc ctgatcacca cacttggcga gatcagtcca gttctgccac
     2221 ctgtatcctt tggtgccact tcagtcttgg ctggaatggc tgtcgcctgc ttcctgactg
     2281 agacccgcaa tgtgccactg gtagaaacta ttgctgcaat ggagagacga gtcaaacaag
     2341 gccgttccaa aagagacaca gaacagaaga gtgaagaaat ttctcttcag cagctggggg
     2401 cttcacccct caaagaaacc attggatccg actacaaaga ccatgacggt gattataaag
     2461 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     2521 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2581 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2641 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2701 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2761 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2821 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2881 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2941 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3001 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3061 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3121 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3181 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3241 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3301 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3361 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3421 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3781 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3841 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4021 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4081 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4201 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4261 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4321 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4381 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4441 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4501 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4561 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4621 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4681 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4741 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4801 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4861 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4921 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4981 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5041 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5101 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5161 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5221 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5281 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5341 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5401 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5461 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5521 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5581 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5641 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5701 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6241 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7561 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     7621 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7681 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7741 agg
//
