LOCUS       V024998                 5613 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024998
VERSION     V024998
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             641..1510
                     /label="CD274(NM_014143)"
                     /note="CD274(NM_014143)"
                     /gene="CD274"
     CDS             1559..2278
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced blue variant of GFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EBFP"
     polyA_signal    2401..2522
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(2529..2984)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3011..3115
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3117..3474
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3509..4303
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4535..4582
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4911..5499
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atgaggatat ttgctgtctt
      661 tatattcatg acctactggc atttgctgaa cgcatttact gtcacggttc ccaaggacct
      721 atatgtggta gagtatggta gcaatatgac aattgaatgc aaattcccag tagaaaaaca
      781 attagacctg gctgcactaa ttgtctattg ggaaatggag gataagaaca ttattcaatt
      841 tgtgcatgga gaggaagacc tgaaggttca gcatagtagc tacagacaga gggcccggct
      901 gttgaaggac cagctctccc tgggaaatgc tgcacttcag atcacagatg tgaaattgca
      961 ggatgcaggg gtgtaccgct gcatgatcag ctatggtggt gccgactaca agcgaattac
     1021 tgtgaaagtc aatgccccat acaacaaaat caaccaaaga attttggttg tggatccagt
     1081 cacctctgaa catgaactga catgtcaggc tgagggctac cccaaggccg aagtcatctg
     1141 gacaagcagt gaccatcaag tcctgagtgg taagaccacc accaccaatt ccaagagaga
     1201 ggagaagctt ttcaatgtga ccagcacact gagaatcaac acaacaacta atgagatttt
     1261 ctactgcact tttaggagat tagatcctga ggaaaaccat acagctgaat tggtcatccc
     1321 agaactacct ctggcacatc ctccaaatga aaggactcac ttggtaattc tgggagccat
     1381 cttattatgc cttggtgtag cactgacatt catcttccgt ttaagaaaag ggagaatgat
     1441 ggatgtgaaa aaatgtggca tccaagatac aaactcaaag aagcaaagtg atacacattt
     1501 ggaggagacg ggtggaggcg gttcaggtgg cggaggttcg gatccaccgg tcgccaccat
     1561 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1621 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1681 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1741 cgtgaccacc ctgacccacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1801 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1861 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1921 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1981 gctggagtac aacttcaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2041 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2101 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2161 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2221 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     2281 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2341 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2401 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2461 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2521 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2581 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2641 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2701 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2761 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2821 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2881 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2941 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3001 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3061 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3121 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3181 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3301 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3421 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3481 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3661 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4021 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4321 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4441 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4501 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4561 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4621 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4681 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4741 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5041 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5581 gattctgtgg ataaccgtat taccgccatg cat
//
