LOCUS       V039226                 7933 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039226
VERSION     V039226
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             706..2589
                     /label="KLHL14(NM_020805)"
                     /note="KLHL14(NM_020805)"
                     /gene="KLHL14"
     CDS             2596..2685
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2709..3297
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3363..3411
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3613..4041
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4055..4384
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4451..5245
                     /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    5421..5542
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5591..5607)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5615..5631
                     /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(5639..5669)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5684..5705
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6752..7612)
                     /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(7613..7717)
                     /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 gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgtc cagatccggg
      721 gacaggacct ccaccttcga ccccagccac agcgacaacc tgctgcacgg cctcaacctg
      781 ctgtggagga agcagctgtt ttgcgacgtg accctgacgg cccagggcca gcagttccat
      841 tgccacaagg ccgtgctggc ctcctgctcg cagtacttcc gatcgctctt ctccagccac
      901 ccccctctcg ggggaggggt cggcggccag gacggcctgg gggcccccaa ggaccagcag
      961 cagccgccgc agcagcagcc gtcacagcag cagcagccgc cgccgcagga ggagcccggg
     1021 actccttctt cctcccccga cgacaagctg ctgaccagcc cccgggccat caacaacctg
     1081 gtgctgcagg gctgctcgtc catcgggctg cgcctggtgc tcgagtacct ctacacggcc
     1141 aacgtgaccc tgtccctgga cacggtggag gaggtgctgt cggtcagcaa gatcctgcac
     1201 atcccccagg tcaccaagct ctgcgtgcag ttcctcaacg accagatctc ggtgcagaac
     1261 tacaagcagg tgtgcaagat cgccgcgctg cacggcctgg aggagaccaa gaagctggcc
     1321 aacaagtacc tggtggagga tgtgctgctg ctcaacttcg aggagatgcg cgccctgctg
     1381 gactcgctgc cgccccccgt ggagtcggag ctggcgctct tccagatgtc cgtgctgtgg
     1441 ctggagcacg accgcgagac ccgcatgcag tatgcgcctg acctcatgaa gcgcctccgc
     1501 ttcgccctca tcccggcccc ggagctggtg gagcgggtcc agtcagtgga tttcatgcga
     1561 accgacccgg tctgccagaa gctgctgctg gacgccatga actaccacct gatgcccttc
     1621 aggcagcact gcaggcagag cctggccagc agaattcgct ctaacaagaa aatgctgtta
     1681 ttggttggag ggctgcctcc tggaccggac cggctcccca gcaatttggt tcagtattac
     1741 gacgatgaaa agaagacatg gaaaatactc acaattatgc catacaacag tgcccaccac
     1801 tgcgttgtgg aggtggaaaa cttcttgttc gtgttgggtg gagaggacca gtggaatccg
     1861 aatggaaaac acagtacaaa ttttgtcagc cgatatgatc ctcgatttaa tagctggatt
     1921 caacttccac ccatgcagga aagaagagcc agtttctatg catgtcggtt ggacaagcat
     1981 ttatacgtaa ttggtggaag gaatgaaact ggctacttgt ccagcgtgga gtgctataac
     2041 ctagaaacga atgaatggcg ctatgtgtcc tctttgccac agcctctggc ggctcatgcg
     2101 ggagcagtgc acaatgggaa aatatacatt tcagggggtg tacacaatgg agaatatgtc
     2161 ccatggctat attgctatga cccagtaatg gatgtctggg ctcgaaaaca agatatgaac
     2221 acaaaacgtg caattcacac tttggctgta atgaatgatc gcttgtatgc aattggagga
     2281 aatcatttga aaggtttctc ccaccttgat gtaatgcttg tggaatgcta tgacccaaaa
     2341 ggtgaccagt ggaatatact ccaaactccc attttggagg gtcgaagtgg ccctggctgt
     2401 gcagtgcttg atgacagcat ttaccttgtg ggaggctaca gctggagtat gggggcctac
     2461 aagtcatcta caatatgcta ttgtccagag aaaggaacct ggacagaact cgaaggagat
     2521 gtagcagaac cgttggcagg ccctgcctgt gtgacagtta ttctgccctc ttgtgtacca
     2581 tacaacaaac tcgaggaaca aaaactaata tcagaggaag atttggaaca gaaactaatc
     2641 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagtg agggcccgat
     2701 atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2761 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2821 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2881 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2941 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3001 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3061 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     3121 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3181 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3241 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     3301 gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3361 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
     3421 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     3481 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     3541 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta
     3601 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3661 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3721 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag
     3781 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3841 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3901 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3961 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt
     4021 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4081 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4141 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4201 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4261 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4321 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4381 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4441 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4501 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4561 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4621 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4681 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4741 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4801 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4861 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4921 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4981 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5041 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5101 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5161 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5221 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag
     5281 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5341 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5401 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5521 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5641 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg
     7801 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     7861 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     7921 gaatctgctt agg
//
