LOCUS       V045482                 6903 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045482
VERSION     V045482
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..1583
                     /label="Arl14ep(NM_001025102)"
                     /note="Arl14ep(NM_001025102)"
                     /gene="Arl14ep"
     CDS             1590..1655
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1679..2267
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2333..2381
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2583..3011
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3025..3354
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3421..4215
                     /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    4391..4512
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4561..4577)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4585..4601
                     /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(4609..4639)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4654..4675
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4963..5551)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5722..6582)
                     /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(6583..6687)
                     /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 ccaccatgga tccgtgttca gttggagtcc
      781 agcttcgtac cacacacgac tgccataaaa ccttctatac tcggcacaca ggtttcaaga
      841 cattgaaaga attgtcatca aatgacatgc ttttacttca gcttagaact ggaatgacac
      901 tttctgggaa taatacaatt tgtctccatc acgtaaaaat ttatattgac agatttgaag
      961 atttgcagaa gtcgtgttgt gacccattta acatacacaa gaagctagcc aaaaagaacc
     1021 tgcacgtcat tgacttagat gatgccactt ttctgagcgc caagtttgga agacagcttg
     1081 tacctggttg gaagctttgc cccaaatgta cccagatcat caatggaagt gtggacgttg
     1141 attcagatga ccgccagaga cggaaacccg actcagatgg aaggactgct aaagctctga
     1201 ggtcactaca gtttacaaac ccagggaagc aaactgaatt tgctccagaa ggcggtaaga
     1261 gggagaaaag aaagctcacc aaagccacaa gcgctgcttc tgacagacaa attattccag
     1321 caaagagtaa ggtgtatgac agccagggcc tgctgatttt cagtggcatg gacctctgcg
     1381 actgcctgga cgaggactgc ttgggatgct tctacgcttg ccccacttgt ggctctacca
     1441 aatgtggggc tgagtgccgc tgcgaccgaa agtggctgta tgagcagata gagattgaag
     1501 gaggagagat aatccataat aaacatgctg gaaaagcata tggtctgtta tctccctgtc
     1561 acccatatga tattttacaa aaaggatccg actacaaaga ccatgacggt gattataaag
     1621 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     1681 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1741 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1801 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     1861 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     1921 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     1981 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2041 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2101 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2161 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2221 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     2281 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     2341 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     2401 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     2461 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     2521 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     2581 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2641 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2701 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     2761 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2821 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2881 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2941 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3001 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3061 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3181 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3241 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3301 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3361 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3421 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3481 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3541 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3601 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3661 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3721 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3781 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3841 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3901 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3961 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4021 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4081 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4141 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4201 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     4261 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4321 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4381 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4441 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4501 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4561 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4621 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4681 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4741 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4801 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5401 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5641 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5701 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5761 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5821 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5881 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5941 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6001 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6061 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6121 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6181 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6241 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6301 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6361 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6421 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6481 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6541 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6601 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6661 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6721 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     6781 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6841 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     6901 agg
//
