LOCUS       sequence-314046-        8449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8449)
  AUTHORS   Baumann C, Korner R, Hofmann K, Nigg EA
  TITLE     PICH, a centromere-associated SNF2 family ATPase, is regulated by 
            Plk1 and required for the spindle checkpoint.
  JOURNAL   Cell. 2007 Jan 12;128(1):101-14.
  PUBMED    17218258
REFERENCE   2  (bases 1 to 8449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "2007-01-12"; volume: "128(1)"; pages: "101-14"
COMMENT     Sequence Label: pEGFP PICH (Nigg CB62)
FEATURES             Location/Qualifiers
     source          1..8449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5177..5298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5214..5233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5268..5287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5305..5760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5442..5463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5654..5673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5787..5891
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5893..6250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5893..6084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5916..5936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6101..6236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6285..7079
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6285..7079
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6339..6358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6949..6968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7267..7286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7311..7358
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7687..8275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8176..8195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct atggaggcat cccgaaggtt tccggaagcc
     1321 gaggccttga gcccagagca ggctgctcat tacctaagat atgtgaaaga ggccaaagaa
     1381 gcaactaaga atggagacct ggaagaagca tttaaacttt tcaatttggc aaaggacatt
     1441 tttcccaatg aaaaagtgct gagcagaatc caaaaaatac aggaagcctt ggaggagttg
     1501 gcagaacagg gagatgatga atttacagat gtgtgcaact ctggcttgct actttatcga
     1561 gaactgcaca accaactctt tgagcaccag aaggaaggca tagctttcct ctatagcctg
     1621 tatagggatg gaagaaaagg tggtatattg gctgatgata tgggattagg gaagactgtt
     1681 caaatcattg ctttcctttc cggtatgttt gatgcatcac ttgtgaatca tgtgctgctg
     1741 atcatgccaa ccaatcttat taacacatgg gtaaaagaat tcatcaagtg gactccagga
     1801 atgagagtca aaacctttca tggtcctagc aaggatgaac ggaccagaaa cctcaatcgg
     1861 attcagcaaa ggaatggtgt tattatcact acataccaaa tgttaatcaa taactggcag
     1921 caactttcaa gctttagggg ccaagagttt gtgtgggact atgtcatcct cgatgaagca
     1981 cataaaataa aaacctcatc tactaagtca gcaatatgtg ctcgtgctat tcctgcaagt
     2041 aatcgcctcc tcctcacagg aaccccaatc cagaataatt tacaagaact atggtcccta
     2101 tttgattttg cttgtcaagg gtccctgctg ggaacattaa aaacttttaa gatggagtat
     2161 gaaaatccta ttactagagc aagagagaag gatgctaccc caggagaaaa agccttggga
     2221 tttaaaatat ctgaaaactt aatggcaatc ataaaaccct attttctcag gaggactaaa
     2281 gaagacgtac agaagaaaaa gtcaagcaac ccagaggcca gacttaatga aaagaatcca
     2341 gatgttgatg ccatttgtga aatgccttcc ctttccagga aaaatgattt aattatttgg
     2401 atacgacttg tgcctttaca agaagaaata tacaggaaat ttgtgtcttt agatcatatc
     2461 aaggagttgc taatggagac gcgctcacct ttggctgagc taggtgtctt aaagaagctg
     2521 tgtgatcatc ctaggctgct gtctgcacgg gcttgttgtt tgctaaatct tgggacattc
     2581 tctgctcaag atggaaatga gggggaagat tccccagatg tggaccatat tgatcaagta
     2641 actgatgaca cattgatgga agaatctgga aaaatgatat tcctaatgga cctacttaag
     2701 aggctgcgag atgagggaca tcaaactctg gtgttttctc aatcgaggca aattctaaac
     2761 atcattgaac gcctcttaaa gaataggcac tttaagacat tgcgaatcga tgggacagtt
     2821 actcatcttt tggaacgaga aaaaagaatt aacttattcc agcaaaataa agattactct
     2881 gtttttctgc ttaccactca agtaggtggt gtcggtttaa cattaactgc agcaactaga
     2941 gtggtcattt ttgaccctag ctggaatcct gcaactgatg ctcaagctgt ggatagagtt
     3001 taccgaattg gacaaaaaga gaatgttgtg gtttataggc taatcacttg tgggactgta
     3061 gaggaaaaaa tatacagaag acaggttttc aaggactcat taataagaca aactactggt
     3121 gaaaaaaaga accctttccg atattttagt aaacaagaat taagagagct ctttacaatc
     3181 gaggatcttc agaactctgt aacccagctg cagcttcagt ctttgcatgc tgctcagagg
     3241 aaatctgata taaaactaga tgaacatatt gcctacctgc agtctttggg gatagctgga
     3301 atctcagacc atgatttgat gtacacatgt gatctgtctg ttaaagaaga gcttgatgtg
     3361 gtagaagaat ctcactatat tcaacaaagg gttcagaaag ctcaattcct cgttgaattc
     3421 gagtctcaaa ataaagagtt cctgatggaa caacaaagaa ctagaaatga gggggcctgg
     3481 ctaagagaac ctgtatttcc ttcttcaaca aagaagaaat gccctaaatt gaataaacca
     3541 cagcctcagc cttcacctct tctaagtact catcatactc aggaagaaga tatcagttcc
     3601 aaaatggcaa gtgtagtcat tgatgatctg cccaaagagg gtgagaaaca agatctctcc
     3661 agtataaagg tgaatgttac caccttgcaa gatggtaaag gtacaggtag tgctgactct
     3721 atagctactt taccaaaggg gtttggaagt gtagaagaac tttgtactaa ctcttcattg
     3781 ggaatggaaa aaagctttgc aactaaaaat gaagctgtac aaaaagagac attacaagag
     3841 gggcctaagc aagaggcact gcaagaggat cctctggaaa gttttaatta tgtacttagc
     3901 aaatcaacca aagctgatat tgggccaaat ttagatcaac taaaggatga tgagatttta
     3961 cgtcattgca atccttggcc cattatttcc ataacaaatg aaagtcaaaa tgcagaatca
     4021 aatgtatcca ttattgaaat agctgatgac ctttcagcat cccatagtgc actgcaggat
     4081 gctcaagcaa gtgaggccaa gttggaagag gaaccttcag catcttcacc acagtatgca
     4141 tgtgatttca atcttttctt ggaagactca gcagacaaca gacaaaactt ttccagtcag
     4201 tctttagagc atgttgagaa agaaaatagc ttgtgtggct ctgcacctaa ttccagagca
     4261 gggtttgtgc atagcaaaac atgtctcagt tgggagtttt ctgagaaaga cgatgaacca
     4321 gaagaagtag tagttaaagc aaaaatcaga agtaaagcta gaaggattgt ttcagatggc
     4381 gaagatgaag atgattcttt taaagatacc tcaagcataa atccattcaa cacatctctc
     4441 tttcaattct catctgtgaa acaatttgat gcttcaactc ccaaaaatga catcagtcca
     4501 ccaggaaggt tcttttcatc tcaaataccc agtagtgtaa ataagtctat gaactctaga
     4561 agatctctgg cttctaggag gtctcttatt aatatggttt tagaccacgt ggaggacatg
     4621 gaggaaagac ttgacgacag cagtgaagca aagggtcctg aagattatcc agaagaaggg
     4681 gtggaggaaa gcagtggcga agcctccaag tatacagaag aggatccttc cggagaaaca
     4741 ctgtcttcag aaaacaagtc cagctggtta atgacgtcta agcctagtgc tctagctcaa
     4801 gagacctctc ttggtgcccc tgagcctttg tctggtgaac agttggttgg ttctccccag
     4861 gataaggcgg cagaggctac aaatgactat gagactcttg taaagcgtgg aaaagaacta
     4921 aaagagtgtg gaaaaatcca ggaggcccta aactgcttag ttaaagcgct tgacataaaa
     4981 agtgcagatc ctgaagttat gctcttgact ttaagtttgt ataagcaact taataacaat
     5041 tgacccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     5101 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5161 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5221 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5281 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5341 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5401 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5461 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5521 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5581 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5641 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5701 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5761 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5821 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5881 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6001 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6181 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6241 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6301 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6361 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6421 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6481 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6541 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6601 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6661 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6721 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6781 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6841 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6901 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6961 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7021 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7081 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7141 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7201 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7261 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7321 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7381 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7441 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7501 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7561 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7621 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7681 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7741 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7801 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7861 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7921 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7981 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8041 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8101 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8161 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8221 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8281 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8341 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     8401 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
