LOCUS       V035071                 6551 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035071
VERSION     V035071
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             634..2460
                     /label="KLHL20(NM_014458)"
                     /note="KLHL20(NM_014458)"
                     /gene="KLHL20"
     CDS             2497..3216
                     /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    3339..3460
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3467..3922)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3949..4053
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4055..4412
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4447..5241
                     /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    5473..5520
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5849..6437
                     /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 caagcttgcc accatggaag gaaagccaat gcgcaggtgt
      661 accaacattc gaccaggaga gactggaatg gatgtaacaa gccgctgcac ccttggagac
      721 cccaacaaac tgccagaagg ggttccccaa cctgcccgca tgccctatat ctcagacaag
      781 caccctcgac aaaccttgga agtgattaac cttctgagaa agcaccggga gctatgtgat
      841 gtggtgctag ttgtgggcgc caagaagata tatgcccatc gagtcatttt gtcagcctgt
      901 agtccctact tccgagctat gtttacagga gaattggcag agagccgtca gacagaagta
      961 gtgatccgag acattgacga gagggctatg gaattactga ttgactttgc gtatacctcc
     1021 cagataacag tagaagaggg caatgttcag actcttctgc cagctgcttg cctcctccag
     1081 ctggcagaaa tacaggaagc ctgctgtgaa ttcttaaaga gacaattaga tccttctaac
     1141 tgcctgggca ttcgggcttt tgctgacaca cattcatgtc gtgagttgct aaggatagca
     1201 gacaagttca cccaacataa ctttcaagag gtaatggaga gtgaagagtt catgttgctt
     1261 ccagccaatc aactcattga tataatatcc agtgatgagc taaacgttcg cagtgaagaa
     1321 caagtgttca atgcagtgat ggcctgggtc aaatacagta ttcaggaaag acgtcctcaa
     1381 ttaccccagg tgctgcagca tgttcgtttg cctttgctta gtcccaagtt cctggtcggc
     1441 acagtaggct ctgatcccct catcaaaagt gatgaagaat gcagagactt ggtagatgag
     1501 gctaaaaact acctcctatt gccgcaagaa cgaccactaa tgcaaggacc aaggacgaga
     1561 ccacggaaac ctatccgatg tggggaagta ctctttgcag ttggtggttg gtgcagtgga
     1621 gatgccattt ccagtgttga acgatatgat ccacagacca atgaatggag aatggtggct
     1681 tcaatgagca aaaggagatg cggagttggg gtcagtgttc ttgatgatct gttatatgca
     1741 gtaggaggcc atgatggatc ctcttatctc aatagtgttg aaaggtatga ccccaaaaca
     1801 aaccagtgga gcagtgatgt ggcccctaca agcacctgca ggacaagtgt tggtgtagca
     1861 gtacttggag gctttcttta tgctgtgggt ggccaggatg gtgtgtcttg cctcaacatt
     1921 gttgagaggt atgatccgaa ggagaacaag tggactcggg tagcttctat gagtaccaga
     1981 agactaggtg tggctgtggc tgtgttagga gggttcttat atgctgtagg tggctctgac
     2041 gggacatctc ctctcaacac agtggaacgt tacaatcctc aggaaaacag atggcacact
     2101 atagccccta tggggacccg gaggaaacac ctaggctgtg cagtatatca ggacatgatc
     2161 tatgctgtag gaggtagaga tgacactaca gagctgagca gtgctgagag atacaacccc
     2221 agaaccaacc agtggtctcc agtggtggcc atgacatcac gccgtagtgg agttggcctg
     2281 gcagtggtca atggacagct catggcagta ggaggttttg atggcacaac atacttgaag
     2341 accatagaag tttttgatcc tgatgccaat acatggaggt tatatggcgg gatgaattac
     2401 cgtcggctag ggggtggcgt aggagttatt aaaatgacac attgtgaatc ccatatttgg
     2461 ggatccggcg gcggagggag tggaggaggg ggaagcatgg tgagcaaggg cgaggagctg
     2521 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2581 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2641 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacccacggc
     2701 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2761 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2821 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2881 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc
     2941 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3001 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3061 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3121 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3181 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     3241 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3301 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3361 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3421 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3481 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3541 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3601 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3661 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3721 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3781 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3841 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3901 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4021 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4081 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4201 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4261 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4321 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4381 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4441 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4501 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4561 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4621 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4681 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4741 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4801 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4861 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4921 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4981 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5041 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5101 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5161 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5221 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5461 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5521 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5581 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5641 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6421 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6481 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6541 ccgccatgca t
//
