LOCUS       V043467                 8318 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043467
VERSION     V043467
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(1..105)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     728..744
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        1440..2618
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             2678..2704
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     protein_bind    3377..3401
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     promoter        3453..3664
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3677..3945
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3978..4772
                     /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="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4889..5010
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6297..6313)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6321..6337
                     /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(6345..6375)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6390..6411
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6699..7287)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7458..8318)
                     /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"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc cacggccact agtattatgc ccagtacatg
     1381 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1441 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     1501 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     1561 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     1621 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     1681 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     1741 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     1801 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     1861 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     1921 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     1981 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     2041 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     2101 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     2161 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     2221 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     2281 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     2341 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     2401 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     2461 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     2521 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     2581 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtacgctagc
     2641 gtttaaactt aagcttggta ccgaattcgc caccatgtac ccatacgatg ttccagatta
     2701 cgctgtttcg aaaggcgagg agctgttcac aggcgtggtg ccaatcctgg tggagctgga
     2761 cggcgacgtg aacggccaca aattcagcgt gagaggcgag ggcgagggcg acgccacaat
     2821 cggcaaactg acactgaaat tcatctgcac aacaggcaaa ctgccagtgc cctggccaac
     2881 actagtgaca acactgacat acggcgtgca gtgcttcagc agatatccgg accacatgaa
     2941 aagacacgac ttcttcaaaa gcgccatgcc agagggctac gtgcaggaga gaacaatcag
     3001 cttcaaagac gacggcaaat acaaaacaag agccgtggtg aaattcgagg gcgacacact
     3061 ggtgaacaga atcgagctga aaggcacaga cttcaaagag gacggcaaca tcctgggcca
     3121 caaactggag tacaacttca acagccacaa cgtgtacatc acagccaaca aacagaaaaa
     3181 cggcatcaaa gccaacttca cagtgagaca caacgtggag gacggcagcg tgcagctcgc
     3241 cgaccactac cagcagaaca caccaatcgg cgacggccca gtgctgctgc cagacaacca
     3301 ctacctgagc acacagacag tgctgagcaa agacccaaac gagaaatccg gactcagatc
     3361 tcgagcttct agatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt agcggccgcg
     3421 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     3481 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
     3541 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     3601 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     3661 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     3721 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     3781 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     3841 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     3901 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     3961 gcctacgcta ggccaccatg gctgaacaag atggattgca cgcaggttct ccggccgctt
     4021 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4081 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4141 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4201 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4261 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4321 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4381 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4441 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4501 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4561 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4621 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4681 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4741 ccttctatcg ccttcttgac gagttcttct gagtcgacaa tcaacctctg gattacaaaa
     4801 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     4861 ctgctttaat gcctttgtat catgcgttaa cttgtttatt gcagcttata atggttacaa
     4921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4981 tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca
     5041 attagtctat cagaagctat ctggtctccc ttccggggga caagacatcc ctgtttaata
     5101 tttaaacagc agtgttccca aactgggttc ttatatccct tgctctggtc aaccaggttg
     5161 cagggtttcc tgtcctcaca ggaacgaagt ccctaaagaa acagtggcag ccaggtttag
     5221 ccccggaatt gactggattc cttttttagg gcccattggt atggcttttt ccccgtatcc
     5281 ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg
     5341 ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg cggggggagc
     5401 gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg gagggacgta
     5461 attacatccc tgggggcttt gggggggggc tgtccctgat atctataaca agaaaatata
     5521 tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt
     5581 aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata
     5641 gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc
     5701 gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata
     5761 tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca
     5821 tcaggatcat atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct
     5881 gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga
     5941 gtttgccgag gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat
     6001 gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg
     6061 ggataccagt tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat
     6121 cagcaacccg aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga
     6181 cagcggtgcg gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca
     6241 gaaatggaca tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca
     6301 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6361 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6421 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6481 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6541 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6601 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6661 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6721 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6781 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6841 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6901 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6961 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7021 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7081 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7141 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7201 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7261 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7321 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7381 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7441 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7501 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7561 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7621 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7681 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7741 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7801 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7861 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7921 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7981 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8041 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8101 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8161 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8221 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8281 aagggaataa gggcgacacg gaaatgttga atactcat
//
