LOCUS       V027831                 7323 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027831
VERSION     V027831
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..2003
                     /label="Lamp2(NM_001290485)"
                     /note="Lamp2(NM_001290485)"
                     /gene="Lamp2"
     CDS             2010..2075
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2099..2687
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2753..2801
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3003..3431
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3445..3774
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3841..4635
                     /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    4811..4932
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4981..4997)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5005..5021
                     /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(5029..5059)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5074..5095
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5383..5971)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6142..7002)
                     /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(7003..7107)
                     /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 ccaccatgtg cctctctccg gttaaaggcg
      781 caaagctcat cctgatcttt ctgttcctag gagccgttca gtccaatgca ttgatagtta
      841 atttgacaga ttcaaagggt acttgccttt atgcagaatg ggagatgaat ttcacaataa
      901 catatgaaac tacaaaccaa accaataaaa ctataaccat tgcagtacct gacaaggcga
      961 cacacgatgg aagcagttgt ggggatgacc ggaatagtgc caaaataatg atacaatttg
     1021 gattcgctgt ctcttgggct gtgaatttta ccaaggaagc atctcattat tcaattcatg
     1081 acatcgtgct ttcctacaac actagtgata gcacagtatt tcctggtgct gtagctaaag
     1141 gagttcatac tgttaaaaat cctgagaatt tcaaagttcc attggatgtc atctttaagt
     1201 gcaatagtgt tttaacttac aacctgactc ctgtcgttca gaaatattgg ggtattcacc
     1261 tgcaagcttt tgtccaaaat ggtacagtga gtaaaaatga acaagtgtgt gaagaagacc
     1321 aaactcccac cactgtggca cccatcattc acaccactgc cccgtcgact acaactacac
     1381 tcactccaac ttcaacaccc actccaactc caactccaac tccaaccgtt ggaaactaca
     1441 gcattagaaa tggcaatact acctgtctgc tggctaccat ggggctgcag ctgaacatca
     1501 ctgaggagaa ggtgcctttc atttttaaca tcaaccctgc cacaaccaac ttcaccggca
     1561 gctgtcaacc tcaaagtgct caacttaggc tgaacaacag ccaaattaag tatcttgact
     1621 ttatctttgc tgtgaaaaat gaaaaacggt tctatctgaa ggaagtgaat gtctacatgt
     1681 atttggctaa tggctcagct ttcaacattt ccaacaagaa ccttagcttc tgggatgccc
     1741 ctctgggaag ttcttatatg tgcaacaaag agcaggtgct ttctgtgtct agagcgtttc
     1801 agatcaacac ctttaaccta aaggtgcaac cttttaatgt gacaaaagga cagtattcta
     1861 cagctgagga atgtgctgct gactctgacc tcaactttct tattcctgtt gcagtgggtg
     1921 tggccttggg cttccttata attgctgtgt ttatatctta catgattgga agacggaaaa
     1981 gtcgtactgg ttatcagtct gtcggatccg actacaaaga ccatgacggt gattataaag
     2041 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     2101 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2161 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2221 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2281 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2341 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2401 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2461 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2521 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2581 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2641 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     2701 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     2761 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     2821 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     2881 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     2941 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3001 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3061 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3121 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3181 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3241 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3301 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3361 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3421 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3481 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3661 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3721 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3781 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3841 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3901 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3961 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4021 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4081 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4141 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4201 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4261 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4321 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4381 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4441 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4501 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4561 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4621 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     4681 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4741 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4801 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4861 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4921 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4981 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5041 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5101 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5821 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7141 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     7201 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7261 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7321 agg
//
