LOCUS       sequence-238438-        5584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP fused to the N-terminus of Mouse Capping Protein 
            Alpha1, using the Clontech vector PEYFP C-1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  AUTHORS   Edwards M, McConnell P, Schafer DA, Cooper JA
  TITLE     CPI motif interaction is necessary for capping protein function in 
            cells.
  JOURNAL   Nat Commun. 2015 Sep 28;6:8415. doi: 10.1038/ncomms9415.
  PUBMED    26412145
REFERENCE   2  (bases 1 to 5584)
  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: "Nat 
            Commun."; date: "2015-09-28"; volume: "6:8415. doi"; pages: " 
            10.1038/ncomms9415"
COMMENT     Sequence Label: pBJ 2358
FEATURES             Location/Qualifiers
     source          1..5584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(716..737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1324..1345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2430..2551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2467..2486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2521..2540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2558..3013)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2695..2716)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2907..2926
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3040..3144
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3146..3503
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3146..3337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3169..3189)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3354..3489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3538..4332
                     /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             3538..4332
                     /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(3592..3611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4202..4221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4520..4539)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4564..4611
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4940..5528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5429..5448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      841 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
      901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1261 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1381 gtacaagtcc ggactcagat ctcgaggaat ggccgacttt gaggatcggg tgtcggatga
     1441 agagaaggtc cgcatagccg ctaaattcat cacccatgca cccccagggg agtttaatga
     1501 agtattcaat gatgtccggc tactgcttaa caatgacaat ctcctcaggg aaggggcagc
     1561 acatgcattt gcccagtata acatggatca gttcacaccc gtgaagatag aaggctacga
     1621 tgaccaggtc ttaattacag agcatggtga ccttggtaac agcagatttt tagatccaag
     1681 aaaccaaatt tcctttaagt ttgatcattt acggaaagaa gcaagtgacc cgcagccaga
     1741 ggatgtggat ggaggcttga agtcttggag ggagtcgtgt gacagtgctc tgagagccta
     1801 tgtgaaagat cattattcca acggcttctg tactgtttac gctaaaacta tagatgggca
     1861 acagaccatt attgcatgta ttgaaagcca ccagtttcaa ccaaaaaact tctggaatgg
     1921 tcgctggaga tcagagtgga agttcaccat tacaccacct tcagcccagg tggttggagt
     1981 actcaagatt caggttcatt attatgaaga tggcaatgtt cagctggtta gtcataaaga
     2041 tgtgcaggac tcagtaactg tttctaatga aatccaaact accaaggagt ttattaaaat
     2101 catagagagt gcagaaaatg agtatcagac agcaattagt gaaaattacc aaactatgtc
     2161 agacaccacg ttcaaggcct tgcgccggca gcttccagtt acccgcacca aaatcgactg
     2221 gaacaagata ctcagttaca agattggcaa agaaatgcag aatgcttaag aattctgcag
     2281 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     2341 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2401 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2521 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2581 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2641 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2701 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2761 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2821 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2881 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2941 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3001 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3061 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3121 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3181 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3241 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3301 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3421 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3481 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3541 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3601 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3661 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     3721 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3781 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3841 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3901 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3961 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4021 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4081 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4141 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4201 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4261 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4321 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4381 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4441 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4501 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4561 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4621 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4681 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4741 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4801 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4861 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4921 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4981 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5041 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5101 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5161 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5221 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5281 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5341 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5401 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5461 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5521 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5581 ctca
//
