LOCUS       V019893                 8273 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019893
VERSION     V019893
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             650..2176
                     /label="LCK(NM_001042771)"
                     /note="LCK(NM_001042771)"
                     /gene="LCK"
     CDS             2204..2920
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             2960..3025
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3049..3637
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3703..3751
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3953..4381
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4395..4724
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4791..5585
                     /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    5761..5882
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5931..5947)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5955..5971
                     /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(5979..6009)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6024..6045
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6333..6921)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7092..7952)
                     /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(7953..8057)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gctgccacca tgggctgtgg
      661 ctgcagctca cacccggaag atgactggat ggaaaacatc gatgtgtgtg agaactgcca
      721 ttatcccata gtcccactgg atggcaaggg cacgctgctc atccgaaatg gctctgaggt
      781 gcgggaccca ctggttacct acgaaggctc caatccgccg gcttccccac tgcaagacaa
      841 cctggttatc gctctgcaca gctatgagcc ctctcacgac ggagatctgg gctttgagaa
      901 gggggaacag ctccgcatcc tggagcagag cggcgagtgg tggaaggcgc agtccctgac
      961 cacgggccag gaaggcttca tccccttcaa ttttgtggcc aaagcgaaca gcctggagcc
     1021 cgaaccctgg ttcttcaaga acctgagccg caaggacgcg gagcggcagc tcctggcgcc
     1081 cgggaacact cacggctcct tcctcatccg ggagagcgag agcaccgcgg gatcgttttc
     1141 actgtcggtc cgggacttcg accagaacca gggagaggtg gtgaaacatt acaagatccg
     1201 taatctggac aacggtggct tctacatctc ccctcgaatc acttttcccg gcctgcatga
     1261 actggtccgc cattacacca atgcttcaga tgggctgtgc acacggttga gccgcccctg
     1321 ccagacccag aagccccaga agccgtggtg ggaggacgag tgggaggttc ccagggagac
     1381 gctgaagctg gtggagcggc tgggggctgg acagttcggg gaggtgtgga tggggtacta
     1441 caacgggcac acgaaggtgg cggtgaagag cctgaagcag ggcagcatgt ccccggacgc
     1501 cttcctggcc gaggccaacc tcatgaagca gctgcaacac cagcggctgg ttcggctcta
     1561 cgctgtggtc acccaggagc ccatctacat catcactgaa tacatggaga atgggagtct
     1621 agtggatttt ctcaagaccc cttcaggcat caagttgacc atcaacaaac tcctggacat
     1681 ggcagcccaa attgcagaag gcatggcatt cattgaagag cggaattata ttcatcgtga
     1741 ccttcgggct gccaacattc tggtgtctga caccctgagc tgcaagattg cagactttgg
     1801 cctagcacgc ctcattgagg acaacgagta cacagccagg gagggggcca agtttcccat
     1861 taagtggaca gcgccagaag ccattaacta cgggacattc accatcaagt cagatgtgtg
     1921 gtcttttggg atcctgctga cggaaattgt cacccacggc cgcatccctt acccagggat
     1981 gaccaacccg gaggtgattc agaacctgga gcgaggctac cgcatggtgc gccctgacaa
     2041 ctgtccagag gagctgtacc aactcatgag gctgtgctgg aaggagcgcc cagaggaccg
     2101 gcccaccttt gactacctgc gcagtgtgct ggaggacttc ttcacggcca cagagggcca
     2161 gtaccagcct cagcctggag gagggggaag cggatccgcc accatggtga gcaagggcga
     2221 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2281 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2341 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
     2401 ctacggcctg aagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2461 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2521 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2581 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2641 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2701 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2761 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     2821 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2881 cgccgccggg atcactctcg gcatggacga gctgtacaag gaattctgca gatatccagc
     2941 acagtggcgg ccgctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     3001 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     3061 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3121 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3181 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3241 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3301 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3361 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3421 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3481 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3541 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3601 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     3661 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3721 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3781 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3841 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     3901 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     3961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4021 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4081 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4141 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4261 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4321 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4381 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4561 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4621 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4681 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4741 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4981 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5041 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5101 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5161 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5221 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5281 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5341 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5401 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5461 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5521 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5581 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     5641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5881 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8101 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     8161 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8221 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
