LOCUS       V021148                 8827 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021148
VERSION     V021148
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..4303
                     /label="MICAL2(NM_014632)"
                     /note="MICAL2(NM_014632)"
                     /gene="MICAL2"
     CDS             4325..4366
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             4376..4393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4422..4646
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4692..5120
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5134..5463
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5530..6324
                     /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    6500..6621
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6670..6686)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6694..6710
                     /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(6718..6748)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6763..6784
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7072..7660)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7831..8691)
                     /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(8692..8796)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgggggaa aacgaggatg agaagcaggc
      961 ccaggcgggg caggtttttg agaactttgt ccaggcatcc acgtgcaaag gtaccctcca
     1021 ggccttcaac attctcacac gacacctgga cctagaccct ctggaccaca gaaactttta
     1081 ttccaagctc aagtccaagg tgaccacctg gaaagccaaa gccctgtggt acaaattgga
     1141 taagcgtggt tcccacaaag agtataagcg agggaagtcg tgcacgaaca ccaagtgtct
     1201 catagttggg ggaggaccct gtggcttgcg cactgccatt gaacttgcct acctgggagc
     1261 caaagtggtc gtggtggaga agagggactc cttctcccgg aacaacgtgc tacacctctg
     1321 gcctttcacc atccatgacc ttcgtggcct gggagccaag aagttctatg ggaagttctg
     1381 tgctggctcc atcgaccata tcagtattcg ccaactacag ctcatcctat tcaaggtggc
     1441 cctgatgctg ggagttgaaa tccatgtgaa tgtggagttc gtgaaggttc tagagcctcc
     1501 tgaagatcaa gaaaatcaaa aaattggctg gcgggcagaa tttctcccta cagaccattc
     1561 tctgtcggag tttgagtttg acgtcatcat tggtgccgat ggccgcagga acaccctgga
     1621 agggttcaga agaaaagaat tccgtgggaa gctggcgatt gccatcaccg ccaacttcat
     1681 aaacagaaac agcacagcgg aagccaaggt ggaagagatt agtggtgtgg ctttcatctt
     1741 caatcagaaa ttttttcagg accttaaaga agaaacaggc atagatcttg agaacattgt
     1801 ttactacaag gactgcaccc actattttgt aatgacagcc aagaagcaga gcctgctcga
     1861 caaaggtgtc atcattaacg actacatcga cacagagatg ctgctgtgtg cggagaacgt
     1921 gaaccaagac aacctgctat cctatgcccg ggaagctgca gactttgcca ccaactacca
     1981 gctgccatcc ttagactttg ccatgaacca ctatgggcag cctgatgtgg ccatgtttga
     2041 ctttacctgc atgtatgcct cagagaacgc ggccctggtg cgggagcggc aggcgcacca
     2101 gctgctcgtg gcccttgtgg gtgacagctt gcttgagcca ttttggccca tgggtacagg
     2161 ctgtgcccgt ggcttcctgg cagcctttga cacggcatgg atggtgaaga gctggaacca
     2221 gggcacccct cccctggagc tgctggctga aagggaaagt ctctaccggc tgttacctca
     2281 gacaaccccg gagaacatca acaagaactt tgagcagtac acgttggacc cagggacacg
     2341 gtacccaaac ctcaactcac actgtgtcag gccccatcag gtgaagcatt tgtatatcac
     2401 taaggagctg gagcactacc ctctcgagag actgggctcg gtgaggagat ctgtcaacct
     2461 ctccaggaag gagtcagata tccggcccag caagctcctg acctggtgcc agcagcagac
     2521 agagggctac cagcatgtca acgtcaccga cctgaccaca tcctggcgca gtgggttggc
     2581 cctgtgtgcc atcatccacc gcttccggcc tgagctcatc aactttgact ctttgaatga
     2641 agatgatgct gtggagaaca accagctcgc atttgatgtg gccgagcgag agtttgggat
     2701 ccctccagtg accacgggca aagagatggc atctgcccag gagcctgaca agctcagcat
     2761 ggtcatgtac ctctccaagt tctacgagct cttccggggc accccactga ggcccgtgga
     2821 ttcttggcgc aaaaactatg gagaaaatgc tgacctcagc ttggccaaat catccatttc
     2881 taataactat ctcaacctca catttccaag gaagaggact ccacgggtgg atggtcaaac
     2941 cggagagaat gacatgaaca aacggagacg gaagggcttc accaacctgg acgagccttc
     3001 aaacttttcc agccgtagct tgggctccaa tcaagagtgt gggagcagta aggaaggtgg
     3061 aaatcagaac aaagtcaagt ccatggcgaa tcagctgctg gccaagtttg aggagagcac
     3121 tcggaacccc tcactcatga agcaggaacg ccgtgtctca gggataggta agccggtcct
     3181 gtgctcttcc tccggccctc ctgttcactc ttgctgcccc aagccggagg aggccacacc
     3241 cagcccatca cctcctctga aaaggcagtt cccctctgtg gtcgtgacgg ggcacgtgct
     3301 cagagagctc aagcaagtgt ctgctggcag tgagtgcctg agcagacctt ggagagccag
     3361 agccaagtct gacctacagc tgggtgggac agaaaatttc gctaccctgc cttctacccg
     3421 cccgagggcg caggctcttt ccggggtgct gtggcggctg cagcaagtgg aggaaaagat
     3481 tctccagaag agggctcaga acttggccaa cagggaattt cacacaaaga acattaagga
     3541 gaaggcggct caccttgcct ccatgtttgg acacggggat ttcccgcaga ataaactact
     3601 ctctaaaggc ctgtctcata ctcatcctcc atctcctccc tctcgccttc cgtctcctga
     3661 tccagctgct tcttcctctc catcaactgt tgactctgct tctcctgcca gaaaggaaaa
     3721 gaagtcacct tcagggttcc attttcatcc cagccatttg agaacagtgc atcctcagct
     3781 gacggtaggg aaagtgtcca gcggaatagg ggctgcagct gaagtcctgg tcaatctgta
     3841 catgaatgat cacagaccta aggcccaggc cacctctcca gacctggaat ctatgcgaaa
     3901 gtcatttccc cttaacctgg gaggcagcga cacgtgttac ttctgtaaga aacgtgtgta
     3961 cgtgatggaa cggctgagcg ccgagggcca cttcttccac cgggagtgtt tccgctgcag
     4021 catctgtgcc accaccttgc gcctggccgc ctacaccttt gactgcgatg aaggcaaatt
     4081 ttactgcaag cctcacttca ttcactgtaa aaccaatagc aaacaacgga agagacgggc
     4141 agagttgaag caacaaagag aggaggaggc aacatggcaa gagcaggaag cccctcggag
     4201 agacactccc accgaaagtt cttgcgcagt ggccgccatt ggcaccctgg aaggcagccc
     4261 cccagttcat ttcagccttc cagtgctaca cccacttctt ggcctagagg gcccgcggtt
     4321 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     4381 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4621 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4681 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5041 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5101 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5401 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5461 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5521 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5581 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5641 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5701 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5761 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5821 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5881 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5941 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6001 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6061 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6121 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6181 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6241 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6301 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6361 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6421 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6481 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6601 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6661 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6721 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6781 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6841 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8821 tgacgtc
//
