LOCUS       sequence-122954-        6167 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 6167)
  AUTHORS   Unsal-Kacmaz K, Mullen TE, Kaufmann WK, Sancar A
  TITLE     Coupling of human circadian and cell cycles by the timeless protein.
  JOURNAL   Mol Cell Biol. 2005 Apr . 25(8):3109-16.
  PUBMED    15798197
REFERENCE   2  (bases 1 to 6167)
  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: "Mol Cell 
            Biol. 2005 Apr . 25(8):3109-16."
COMMENT     Sequence Label: peGFP-Chk1
FEATURES             Location/Qualifiers
     source          1..6167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2047..2099
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2107..2116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2113..2115,2116..2118,2119..2832)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2113..2115,2116..2118,2119..2829)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2158..2179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2419..2438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2766..2787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2955..3076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2992..3011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3046..3065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3083..3538)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3220..3241)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3432..3451
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3565..3669
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3671..4028
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3671..3862
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3694..3714)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3879..4014
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4063..4857
                     /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             4063..4857
                     /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(4117..4136)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4727..4746
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5045..5064)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5089..5136
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5465..6053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5954..5973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 atctcgagat ggcagtgccc tttgtggaag actgggactt ggtgcaaacc
      661 ctgggagaag gtgcctatgg agaagttcaa cttgctgtga atagagtaac tgaagaagca
      721 gtcgcagtga agattgtaga tatgaagcgt gccgtagact gtccagaaaa tattaagaaa
      781 gagatctgta tcaataaaat gctaaatcat gaaaatgtag taaaattcta tggtcacagg
      841 agagaaggca atatccaata tttatttctg gagtactgta gtggaggaga gctttttgac
      901 agaatagagc cagacatagg catgcctgaa ccagatgctc agagattctt ccatcaactc
      961 atggcagggg tggtttatct gcatggtatt ggaataactc acagggctat taaaccagaa
     1021 aatcttctgt tggatgaaag ggataacctc aaaatctcag actttggctt ggcaacagta
     1081 tttcggtata ataatcgtga gcgtttgttg aacaagatgt gtggtacttt accatatgtt
     1141 gctccagaac ttctgaagag aagagaattt catgcagaac cagttgatgt ttggtcctgt
     1201 ggaatagtac ttactgcaat gctcgctgga gaattgccat gggaccaacc cagtgacagc
     1261 tgtcaggagt attctgactg gaaagaaaaa aaaacatacc tcaacccttg gaaaaaaatc
     1321 gattctgctc ctctagctct gctgcataaa atcttagttg agaatccatc agcaagaatt
     1381 accattccag acatcaaaaa agatagatgg tacaacaaac cccccaagaa aggggcaaaa
     1441 aggccccgag tcacttcagg tggtgtgtca gagtctccca gtggattttc taagcacatt
     1501 caatccaatt tggacttctc tccagtaaac agtgcttcta gtgaagaaaa tgtgaagtac
     1561 tccagttctc agccagaacc ccgcacaggt ctttccttat gggataccag cccctcatac
     1621 attgataaat tggtacaagg gatcagcttt tcccagccca catgtcctga tcatatgctt
     1681 ttgaatagtc agttacttgg caccccagga tcctcacaga acccctggca gcggttggtc
     1741 aaaagaatga cacgattctt taccaaattg gatgcagaca aatcttatca atgcctgaaa
     1801 gagacttgtg agaagttggg ctatcaatgg aagaaaagtt gtatgaatca ggttactata
     1861 tcaacaactg ataggagaaa caataaactc attttcaaag tgaatttgtt agaaatggat
     1921 gataaaatat tggttgactt ccggctttct aagggtgatg gattggagtt caagagacac
     1981 ttcctgaaga ttaaagggaa gctgattgat attgtgagca gccagaaggt ttggcttcct
     2041 gccacactcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     2101 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2161 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2221 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2281 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2341 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2401 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2461 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2521 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2581 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2641 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2701 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2761 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2821 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     2881 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2941 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3061 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3121 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3181 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3241 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3301 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3361 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3421 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3481 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3661 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3721 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3781 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3901 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3961 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4021 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4561 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4861 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4921 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4981 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5041 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5101 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5161 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5221 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5281 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5341 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5401 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5461 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5521 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5581 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5641 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5701 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5761 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5821 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5881 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5941 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6001 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6061 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6121 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
