LOCUS       V033681                 7305 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033681
VERSION     V033681
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..1985
                     /label="LAMP2(NM_002294)"
                     /note="LAMP2(NM_002294)"
                     /gene="LAMP2"
     CDS             1992..2057
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2081..2669
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2735..2783
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2985..3413
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3427..3756
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3823..4617
                     /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    4793..4914
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4963..4979)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4987..5003
                     /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(5011..5041)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5056..5077
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5365..5953)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6124..6984)
                     /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(6985..7089)
                     /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 ccaccatggt gtgcttccgc ctcttcccgg
      781 ttccgggctc agggctcgtt ctggtctgcc tagtcctggg agctgtgcgg tcttatgcat
      841 tggaacttaa tttgacagat tcagaaaatg ccacttgcct ttatgcaaaa tggcagatga
      901 atttcacagt acgctatgaa actacaaata aaacttataa aactgtaacc atttcagacc
      961 atggcactgt gacatataat ggaagcattt gtggggatga tcagaatggt cccaaaatag
     1021 cagtgcagtt cggacctggc ttttcctgga ttgcgaattt taccaaggca gcatctactt
     1081 attcaattga cagcgtctca ttttcctaca acactggtga taacacaaca tttcctgatg
     1141 ctgaagataa aggaattctt actgttgatg aacttttggc catcagaatt ccattgaatg
     1201 acctttttag atgcaatagt ttatcaactt tggaaaagaa tgatgttgtc caacactact
     1261 gggatgttct tgtacaagct tttgtccaaa atggcacagt gagcacaaat gagttcctgt
     1321 gtgataaaga caaaacttca acagtggcac ccaccataca caccactgtg ccatctccta
     1381 ctacaacacc tactccaaag gaaaaaccag aagctggaac ctattcagtt aataatggca
     1441 atgatacttg tctgctggct accatggggc tgcagctgaa catcactcag gataaggttg
     1501 cttcagttat taacatcaac cccaatacaa ctcactccac aggcagctgc cgttctcaca
     1561 ctgctctact tagactcaat agcagcacca ttaagtatct agactttgtc tttgctgtga
     1621 aaaatgaaaa ccgattttat ctgaaggaag tgaacatcag catgtatttg gttaatggct
     1681 ccgttttcag cattgcaaat aacaatctca gctactggga tgcccccctg ggaagttctt
     1741 atatgtgcaa caaagagcag actgtttcag tgtctggagc atttcagata aatacctttg
     1801 atctaagggt tcagcctttc aatgtgacac aaggaaagta ttctacagct caagactgca
     1861 gtgcagatga cgacaacttc cttgtgccca tagcggtggg agctgccttg gcaggagtac
     1921 ttattctagt gttgctggct tattttattg gtctcaagca ccatcatgct ggatatgagc
     1981 aatttggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     2041 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     2101 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2161 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2221 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2281 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2341 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2401 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2461 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2521 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2581 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2641 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     2701 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     2761 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     2821 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     2881 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     2941 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3121 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3781 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4321 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4621 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5161 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5881 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5941 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6001 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6061 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7081 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7141 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7201 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7261 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
