LOCUS       V037396                 9091 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037396
VERSION     V037396
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..3903
                     /label="CSE1L(NM_001316)"
                     /note="CSE1L(NM_001316)"
                     /gene="CSE1L"
     CDS             3941..4660
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4691..4915
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4961..5389
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5403..5732
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5799..6593
                     /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    6767..6888
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6937..6953)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6961..6977
                     /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(6985..7015)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7030..7051
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7339..7924)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8095..8955)
                     /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(8956..9060)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggaactca gcgatgcaaa tctgcaaaca
     1021 ctaacagaat atttaaagaa aacacttgat cctgatcctg ccatccgacg tccagctgag
     1081 aaatttcttg aatctgttga aggaaatcag aattatccac tgttgctttt gacattactg
     1141 gagaagtccc aggataatgt tatcaaagta tgtgcttcag taacattcaa aaactatatt
     1201 aaaaggaact ggagaattgt tgaagatgaa ccaaacaaaa tttgtgaagc cgatcgagtg
     1261 gccattaaag ccaacatagt gcacttgatg cttagcagcc cagagcaaat tcagaagcag
     1321 ttaagtgatg caattagcat tattggcaga gaagattttc cacagaaatg gcctgacttg
     1381 ctgacagaaa tggtgaatcg ctttcagagt ggagatttcc atgttattaa tggagtcctc
     1441 cgtacagcac attcattatt taaaagatac cgtcatgaat ttaagtcaaa cgagttatgg
     1501 actgaaatta agcttgttct ggatgccttt gctttgcctt tgactaatct ttttaaggcc
     1561 actattgaac tctgcagtac ccatgcaaat gatgcctctg ccctgaggat tctgttttct
     1621 tccctgatcc tgatctcaaa attgttctat agtttaaact ttcaggatct ccctgaattt
     1681 tttgaagata atatggaaac ttggatgaat aattttcata ctctcttaac attggataat
     1741 aagcttttac aaactgatga tgaagaggaa gccggcttat tggagctctt aaaatcccag
     1801 atttgtgata atgccgcact ctatgcacaa aagtacgatg aagaattcca gcgatacctg
     1861 cctcgttttg ttacagccat ctggaattta ctagttacaa cgggtcaaga ggttaaatat
     1921 gatttgttgg taagtaatgc aattcaattt ctggcttcag tttgtgagag acctcattat
     1981 aagaatctat ttgaggacca gaacacgctg acaagtatct gtgaaaaggt tattgtgcct
     2041 aacatggaat ttagagctgc tgatgaagaa gcatttgaag ataattctga ggagtacata
     2101 aggagagatt tggaaggatc tgatattgat actagacgca gggctgcttg tgatctggta
     2161 cgaggattat gcaagttttt tgagggacct gtgacaggaa tcttctctgg ttatgttaat
     2221 tccatgctgc aggaatacgc aaaaaatcca tctgtcaact ggaaacacaa agatgcagcc
     2281 atctacctag tgacatcttt ggcatcaaaa gcccaaacac agaagcatgg aattacacaa
     2341 gcaaatgaac ttgtaaacct aactgagttc tttgtgaatc acatcctccc tgatttaaaa
     2401 tcagctaatg tgaatgaatt tcctgtcctt aaagctgacg gtatcaaata tattatgatt
     2461 tttagaaatc aagtgccaaa agaacatctt ttagtctcga ttcctctctt gattaatcat
     2521 cttcaagctg aaagtattgt tgttcatact tacgcagctc atgctcttga acggctcttt
     2581 actatgcgag ggcctaacaa tgccactctc tttacagctg cagaaatcgc accgtttgtt
     2641 gagattctgc taacaaacct tttcaaagct ctcacacttc ctggctcttc agaaaatgaa
     2701 tatattatga aagctatcat gagaagtttt tctctcctac aagaagccat aatcccctac
     2761 atccctactc tcatcactca gcttacacag aagctattag ctgttagtaa gaacccaagc
     2821 aaacctcact ttaatcacta catgtttgaa gcaatatgtt tatccataag aataacttgc
     2881 aaagctaacc ctgctgctgt tgtaaatttt gaggaggctt tgtttttggt gtttactgaa
     2941 atcttacaaa atgatgtgca agaatttatt ccatacgtct ttcaagtgat gtctttgctt
     3001 ctggaaacac acaaaaatga catcccgtct tcctatatgg ccttatttcc tcatctcctt
     3061 cagccagtgc tttgggaaag aacaggaaat attcctgctc tagtgaggct tcttcaagca
     3121 ttcttagaac gcggttcaaa cacaatagca agtgctgcag ctgacaaaat tcctgggtta
     3181 ctaggtgtct ttcagaagct gattgcatcc aaagcaaatg accaccaagg tttttatctt
     3241 ctaaacagta taatagagca catgcctcct gaatcagttg accaatatag gaaacaaatc
     3301 ttcattctgc tattccagag acttcagaat tccaaaacaa ccaagtttat caagagtttt
     3361 ttagtcttta ttaatttgta ttgcataaaa tatggggcac tagcactaca agaaatattt
     3421 gatggtatac aaccaaaaat gtttggaatg gttttggaaa aaattattat tcctgaaatt
     3481 cagaaggtat ctggaaatgt agagaaaaag atctgtgcgg ttggcataac caaattacta
     3541 acagaatgtc ccccaatgat ggacactgag tataccaaac tgtggactcc attattacag
     3601 tctttgattg gtctttttga gttacccgaa gatgatacca ttcctgatga ggaacatttt
     3661 attgacatag aagatacacc aggatatcag actgccttct cacagttggc atttgctggg
     3721 aaaaaagagc atgatcctgt aggtcaaatg gtgaataacc ccaaaattca cctggcacag
     3781 tcacttcaca agttgtctac cgcctgtcca ggaagggttc catcaatggt gagcaccagc
     3841 ctgaatgcag aagcgctcca gtatctccaa gggtaccttc aggcagccag tgtgacactg
     3901 cttctcgagt ggcggtggag gaagcggcgg aggtggatct atggtgagca agggcgagga
     3961 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     4021 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     4081 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     4141 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4201 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4261 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4321 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4381 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4441 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4501 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4561 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4621 cgccgggatc actctcggca tggacgagct gtacaagtag ggcccgttta aacccgctga
     4681 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4741 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4801 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4861 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4921 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4981 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5041 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5101 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5161 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5221 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5281 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5341 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5401 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5461 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5521 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5641 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5701 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5761 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5821 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5881 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5941 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6001 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6061 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6121 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6181 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6241 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6301 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6361 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6421 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6481 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6541 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6601 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6661 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6721 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6781 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6841 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6901 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6961 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7021 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7081 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7141 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7861 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7981 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8041 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8101 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8161 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8221 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8281 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8341 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8401 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8461 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8521 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8581 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8641 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8701 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8761 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8821 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8881 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8941 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9061 cacatttccc cgaaaagtgc cacctgacgt c
//
