LOCUS       V025633                 7885 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025633
VERSION     V025633
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3321
                     /label="AFG3L2(NM_006796)"
                     /note="AFG3L2(NM_006796)"
                     /gene="AFG3L2"
     CDS             3328..3417
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3452..3676
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3722..4150
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4164..4493
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4560..5354
                     /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    5528..5649
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5698..5714)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5722..5738
                     /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(5746..5776)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5791..5812
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6100..6685)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6856..7716)
                     /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(7717..7821)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcgcacc gctgtttgcg gctgtggggc
      961 cggggcggct gctggccccg cggcctacag cagctcctcg tgcctggcgg cgtgggcccg
     1021 ggcgagcagc cctgcctccg gacgctttac cgatttgtta caactcaagc aagggccagc
     1081 agaaattctc ttttgacaga tataattgct gcttatcaaa gattctgttc tcgaccccca
     1141 aaaggatttg aaaaatactt tcctaatgga aaaaatggaa aaaaagctag tgaacctaaa
     1201 gaagttatgg gagagaaaaa agaatcaaag ccagctgcta ccacacgctc ttctggagga
     1261 ggaggtggtg gcggtggaaa acgaggtggc aagaaagatg attctcactg gtggtccagg
     1321 tttcagaagg gtgacattcc atgggacgac aaggatttca ggatgttctt cctctggact
     1381 gctctgttct ggggtggagt catgttttac ttgctgctca agagatccgg gagagaaatc
     1441 acttggaagg actttgtcaa taactatctt tcaaaaggag tagtagacag attggaagtc
     1501 gtcaacaagc gttttgttcg agtgaccttt acaccaggaa aaactcctgt tgatgggcaa
     1561 tacgtttggt ttaatattgg cagtgtggac acctttgaac ggaatctgga aactttacag
     1621 caggaattgg gcatagaagg agaaaatcgg gtgcctgttg tctacattgc tgaaagtgat
     1681 ggctcttttc tgctgagcat gctgcctacg gtgctcatca tcgccttctt gctctacacc
     1741 atcagaagag ggcctgctgg cattggccgg acaggccgag ggatgggcgg actcttcagt
     1801 gtcggagaaa ccactgccaa ggtcttaaag gatgaaattg atgtgaagtt caaagatgtg
     1861 gctggctgtg aggaggccaa gctagagatc atggaatttg tgaatttctt gaaaaaccca
     1921 aagcagtatc aagacctagg agcaaaaatc ccaaagggtg ccattctcac tggtcctcca
     1981 ggcactggga agacgctgct agctaaggcc acagccggag aagccaatgt ccccttcatc
     2041 accgttagtg gatctgagtt tttggagatg ttcgttggtg tgggccctgc tagagtccga
     2101 gacttatttg cccttgctcg gaagaatgcc ccttgcatcc tcttcatcga tgaaatcgat
     2161 gcggtgggaa ggaagagagg aagaggcaac tttggagggc agagtgagca ggagaacaca
     2221 ctcaaccagc tgctggtgga gatggatggt tttaatacaa caacaaatgt cgtcattttg
     2281 gccggcacca atcgaccaga tatcctggac cccgcgctgc ttaggccggg gcgtttcgac
     2341 aggcagatct ttattggacc accagacata aaaggaagag cttctatttt caaagttcat
     2401 ctccgaccgc taaaactgga cagtaccctg gagaaggata aattggcaag aaaactggca
     2461 tctttaactc cagggttttc aggtgctgat gttgctaatg tctgtaatga agctgcgttg
     2521 attgctgcaa ggcatctgtc agattccata aatcagaaac actttgaaca ggcaattgaa
     2581 cgagtgattg gtggcttaga gaagaaaacg caggttctgc agcctgagga gaagaagact
     2641 gtggcatacc acgaagcagg ccatgcggtt gccggctggt atctggagca cgcagacccg
     2701 cttttaaagg tatccatcat cccacgtggc aaaggactag gttatgctca gtatttacca
     2761 aaagaacaat acctctatac caaagagcag ctcttggata ggatgtgtat gactttaggt
     2821 ggtcgagtct ctgaagaaat cttctttgga agaattacaa ctggtgctca agatgacttg
     2881 agaaaagtaa ctcagagtgc atatgcccaa attgttcagt ttggcatgaa tgaaaaggtt
     2941 gggcaaatct cctttgacct cccacgtcag ggggacatgg tattggagaa accttacagt
     3001 gaagccactg caagattgat agatgatgaa gtacgaatac ttattaatga tgcttataaa
     3061 agaacagtag ctcttctcac agaaaagaaa gctgacgtgg agaaggttgc tcttctgttg
     3121 ttagaaaaag aagtattaga taagaatgat atggttgaac ttttgggccc cagaccattt
     3181 gcggaaaaat ctacctatga agaatttgtg gaaggcactg gcagcttgga tgaggacacc
     3241 tcacttccag aaggccttaa ggactggaac aaggagcggg aaaaggagaa agaggagccc
     3301 ccgggtgaga aagttgccaa cctcgaggaa caaaaactaa tatcagagga agatttggaa
     3361 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatga
     3421 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3481 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3541 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3601 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3661 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3721 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4081 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4201 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4381 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4441 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4501 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4561 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4621 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4681 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4741 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4801 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4861 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4921 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4981 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5041 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5101 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5161 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5221 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5281 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5341 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5401 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5461 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5521 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5641 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5701 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5761 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5821 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7861 gggagatctc ccgatcccct atggt
//
