LOCUS       sequence-221149-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  TITLE     Johannes Schmid plasmids
REFERENCE   2  (bases 1 to 7066)
  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"
COMMENT     Sequence Label: Cul1-EYFP
FEATURES             Location/Qualifiers
     source          1..7066
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3854..3975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3891..3910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3945..3964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3982..4437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4119..4140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4331..4350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4464..4568
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4570..4927
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4570..4761
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4593..4613)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4778..4913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4962..5756
                     /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             4962..5756
                     /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(5016..5035)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5626..5645
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5944..5963)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5988..6035
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6364..6952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6853..6872
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atccactagt aacggccgcc agtgtgctgg aattctgcag
     1381 atatcactgt cgtcaacccg gagccagaac ccccacggcc tgaagcagat tggcctggac
     1441 cagatctggg acgacctcag agccggcatc cagcaggtgt acacacggca gagcatggcc
     1501 aagtccagat atatggagct ctacactcat gtttataact actgtactag tgttcaccag
     1561 tcaaaccaag cacgaggagc tggagttcct ccttctaagt cgaaaaaggg gcagacacct
     1621 ggaggagctc agtttgttgg cctggaatta tataaacgac ttaaggaatt tttgaagaat
     1681 tacttgacaa atcttcttaa ggatggagaa gatttgatgg atgagagtgt actgaaattc
     1741 tacactcaac aatgggaaga ttatcgattt tcaagcaaag tgctgaatgg aatttgtgcc
     1801 tacctcaata gacattgggt tcgccgtgaa tgtgacgaag gacgaaaagg aatatatgaa
     1861 atctattcgc ttgcattggt gacttggaga gactgtctgt tcaggccact gaataaacag
     1921 gtaacaaatg ctgttttaaa gctgattgaa aaggaaagga atggtgaaac catcaataca
     1981 agattgatta gtggagttgt acagtcttac gtggaattgg ggctgaatga agatgatgca
     2041 tttgcaaagg gccctacgtt aacagtgtat aaagaatcct ttgaatctca atttttggct
     2101 gacacagaga gattttatac cagagagagt actgaattct tgcagcagaa cccagttact
     2161 gaatatatga aaaaggcaga ggctcgtctg cttgaggaac aacgaagagt tcaggtttac
     2221 cttcatgaaa gcacacaaga tgaattagca aggaaatgtg aacaagtcct cattgaaaaa
     2281 cacttggaaa ttttccacac agaatttcag aatttattgg atgctgacaa aaatgaagat
     2341 ttgggacgca tgtataatct tgtatctaga atccaggatg gcctaggaga attgaaaaaa
     2401 ctgttggaga cacacattca taatcagggt cttgcagcca ttgaaaagtg tggagaagct
     2461 gctttaaatg accccaaaat gtatgtacag acagtgcttg atgttcataa aaaatacaat
     2521 gccctggtaa tgtctgcatt caacaatgac gctggctttg tggctgctct tgataaggct
     2581 tgtggtcgct tcataaacaa caacgcggtt accaagatgg cccaatcatc cagtaaatcc
     2641 cctgagttgc tggctcgata ctgtgactcc ttgttgaaga aaagttccaa gaacccagag
     2701 gaggcagaac tagaagacac actcaatcaa gtgatggttg tcttcaagta catagaagac
     2761 aaagacgtat ttcagaagtt ctatgcgaag atgctcgcca agaggctcgt ccaccagaac
     2821 agtgcaagtg acgatgccga agccagcatg atctccaagt taaagcaagc ttgcgggttc
     2881 gagtacacct ctaaacttca gcgcatgttt caagacattg gcgtgagcaa agatctgaac
     2941 gagcaattca aaaagcactt gacaaactca gaacccctag acttggattt cagcattcaa
     3001 gtgctgagct ccgggtcctg gcccttccag cagtcttgta catttgcctt gccgtcagag
     3061 ttggaacgta gttatcagcg attcacagct ttctacgcca gccgccacag tggccgaaaa
     3121 ttgacgtggt tatatcagtt gtctaaagga gaattggtaa ctaactgctt caaaaacaga
     3181 tatactttgc aggcgtcgac attccagatg gctatcctgc ttcagtacaa cacggaagat
     3241 gcctacactg tgcagcagct gaccgacagc actcaaatta aaatggacat tttggcgcaa
     3301 gttttacaga ttttattaaa gtcgaagcta ttggtcttgg aagatgaaaa tgcaaatgtt
     3361 gatgaggtgg aattgaagcc agatacctta ataaaattat atcttggtta taaaaataag
     3421 aaattaaggg ttaacatcaa tgtgccaatg aaaaccgaac agaagcagga acaagaaacc
     3481 acacacaaaa acatcgagga agaccgcaaa ctactgattc aggcggccat cgtgagaatc
     3541 atgaagatga ggaaggttct gaaacaccag cagttacttg gcgaggtcct cactcagctg
     3601 tcctccaggt tcaaacctcg agtccctgtg atcaagaaat gcattgacat tctaattgag
     3661 aaagaatatt tggagcgagt ggatggtgaa aaggacacct acagttactt ggcttaagcg
     3721 gccgctcgag catgcatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3781 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3841 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3901 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3961 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4021 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4081 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4141 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4201 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4261 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4321 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4381 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4441 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4501 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4561 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4681 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4741 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4801 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4861 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4921 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4981 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5041 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5101 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5161 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5221 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5281 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5341 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5401 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5461 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5521 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5581 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5641 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5701 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5761 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5821 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5881 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5941 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6001 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6061 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6121 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6181 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6301 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6481 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6961 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7021 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
