LOCUS       V021735                 7484 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021735
VERSION     V021735
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             941..2974
                     /label="ACADVL(NM_001270447)"
                     /note="ACADVL(NM_001270447)"
                     /gene="ACADVL"
     CDS             2981..3046
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3084..3308
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3354..3782
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3796..4125
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4192..4986
                     /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    5160..5281
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5330..5346)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5354..5370
                     /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(5378..5408)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5423..5444
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5732..6317)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6488..7348)
                     /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(7349..7453)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgttggggg gcctggccgc
      961 ggcggcggga acccggataa tgggaaagga gatagaagca gaagcacaga ggcccctgag
     1021 gcaaacatgg agacctggcc agccaccagc gatgacagca aagacgatga gctcgcggct
     1081 cacggcgctc ctggggcagc cccggcccgg ccctgcccgg cggccctatg ccgggggtgc
     1141 cgctcagctg gctctggaca agtcagattc ccacccctct gacgctctga ccaggaaaaa
     1201 accggccaag gcggaatcta agtcctttgc tgtgggaatg ttcaaaggcc agctcaccac
     1261 agatcaggtg ttcccatacc cgtccgtgct caacgaagag cagacacagt ttcttaaaga
     1321 gctggtggag cctgtgtccc gtttcttcga ggaagtgaac gatcccgcca agaatgacgc
     1381 tctggagatg gtggaggaga ccacttggca gggcctcaag gagctggggg cctttggtct
     1441 gcaagtgccc agtgagctgg gtggtgtggg cctttgcaac acccagtacg cccgtttggt
     1501 ggagatcgtg ggcatgcatg accttggcgt gggcattacc ctgggggccc atcagagcat
     1561 cggtttcaaa ggcatcctgc tctttggcac aaaggcccag aaagaaaaat acctccccaa
     1621 gctggcatct ggggagactg tggccgcttt ctgtctaacc gagccctcaa gcgggtcaga
     1681 tgcagcctcc atccgaacct ctgctgtgcc cagcccctgt ggaaaatact ataccctcaa
     1741 tggaagcaag ctttggatca gtaatggggg cctagcagac atcttcacgg tctttgccaa
     1801 gacaccagtt acagatccag ccacaggagc cgtgaaggag aagatcacag cttttgtggt
     1861 ggagaggggc ttcgggggca ttacccatgg gccccctgag aagaagatgg gcatcaaggc
     1921 ttcaaacaca gcagaggtgt tctttgatgg agtacgggtg ccatcggaga acgtgctggg
     1981 tgaggttggg agtggcttca aggttgccat gcacatcctc aacaatggaa ggtttggcat
     2041 ggctgcggcc ctggcaggta ccatgagagg catcattgct aaggcggtag atcatgccac
     2101 taatcgtacc cagtttgggg agaaaattca caactttggg ctgatccagg agaagctggc
     2161 acggatggtt atgctgcagt atgtaactga gtccatggct tacatggtga gtgctaacat
     2221 ggaccaggga gccacggact tccagataga ggccgccatc agcaaaatct ttggctcgga
     2281 ggcagcctgg aaggtgacag atgaatgcat ccaaatcatg gggggtatgg gcttcatgaa
     2341 ggaacctgga gtagagcgtg tgctccgaga tcttcgcatc ttccggatct ttgaggggac
     2401 aaatgacatt cttcggctgt ttgtggctct gcagggctgt atggacaaag gaaaggagct
     2461 ctctgggctt ggcagtgctc taaagaatcc ctttgggaat gctggcctcc tgctaggaga
     2521 ggcaggcaaa cagctgaggc ggcgggcagg gctgggcagc ggcctgagtc tcagcggact
     2581 tgtccacccg gagttgagtc ggagtggcga gctggcagta cgggctctgg agcagtttgc
     2641 cactgtggtg gaggccaagc tgataaaaca caagaagggg attgtcaatg aacagtttct
     2701 gctgcagcgg ctggcagacg gggccatcga cctctatgcc atggtggtgg ttctctcgag
     2761 ggcctcaaga tccctgagtg agggccaccc cacggcccag catgagaaaa tgctctgtga
     2821 cacctggtgt atcgaggctg cagctcggat ccgagagggc atggccgccc tgcagtctga
     2881 cccctggcag caagagctct accgcaactt caaaagcatc tccaaggcct tggtggagcg
     2941 gggtggtgtg gtcaccagca acccacttgg cttcctcgag gactacaaag accatgacgg
     3001 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     3061 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3121 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3181 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3241 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3301 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3361 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3421 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3481 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3541 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3601 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3661 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3721 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3781 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3961 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4021 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4081 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4141 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4201 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4261 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4321 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4381 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4441 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4501 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4561 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4621 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4681 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4741 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4801 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4861 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4921 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4981 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5041 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5101 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5281 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6241 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6421 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6481 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6541 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6601 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6661 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6721 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6781 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6841 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6901 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6961 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7021 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7081 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7141 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7201 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7261 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7321 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7381 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7441 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
