LOCUS       sequence-242698-        5700 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 5700)
  AUTHORS   Trinkle-Mulcahy L, Sleeman JE, Lamond AI
  TITLE     Dynamic targeting of protein phosphatase 1 within the nuclei of 
            living mammalian cells.
  JOURNAL   J Cell Sci. 2001 Dec;114(Pt 23):4219-28.
  PUBMED    11739654
REFERENCE   2  (bases 1 to 5700)
  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: "J Cell 
            Sci."; date: "2001-12"; volume: "114(Pt 23)"; pages: "4219-28"
COMMENT     Sequence Label: pEGFP(C1)-PP1gamma
FEATURES             Location/Qualifiers
     source          1..5700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(664..666,667..669,670..1380)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(664..666,667..669,670..1380)
                     /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(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2539..2660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2576..2595)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2630..2649
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2667..3122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2804..2825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3016..3035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3149..3253
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3255..3612
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3255..3446
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3278..3298)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3463..3598
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3647..4441
                     /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             3647..4441
                     /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(3701..3720)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4311..4330
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4629..4648)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4673..4720
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5049..5637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5538..5557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      661 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 tccggactca gatctcgagc tcaagcttcg aattcgatgg cggatttaga taaactcaac
     1441 atcgacagca ttatccaacg gctgctggaa gtgagagggt ccaagcctgg taagaatgtc
     1501 cagcttcagg agaatgaaat cagaggactg tgcttaaagt ctcgtgaaat ctttctcagt
     1561 cagcctatcc tactagaact tgaagcacca ctcaaaatat gtggtgacat ccatggacaa
     1621 tactatgatt tgctgcgact ttttgagtac ggtggtttcc caccagaaag caactacctg
     1681 tttcttgggg actatgtgga caggggaaag cagtcattgg agacgatctg cctcttactg
     1741 gcctacaaaa taaaatatcc tgagaatttt tttcttctca gagggaacca tgaatgtgcc
     1801 agcatcaaca gaatttatgg attttatgat gaatgtaaaa gaagatacaa cattaaacta
     1861 tggaaaactt tcacagactg ttttaactgt ttaccgatag cagccatcgt ggatgagaag
     1921 atattctgtt gtcatggagg tttatcacca gatcttcaat ctatggagca gattcggcga
     1981 attatgcgac caactgatgt accagatcaa ggtcttcttt gtgatctttt gtggtctgac
     2041 cccgataaag atgtcttagg ctggggtgaa aatgacagag gagtgtcctt cacatttggt
     2101 gcagaagtgg ttgcaaaatt tctccataag catgatttgg atcttatatg tagagcccat
     2161 caggtggttg aagatggata tgaatttttt gcaaagaggc agttggtcac tctgttttct
     2221 gcgcccaatt attgcggaga gtttgacaat gcaggtgcca tgatgagtgt ggatgaaaca
     2281 ctaatgtgtt cttttcagat tttaaagcct gcagagaaaa agaagccaaa tgccacgaga
     2341 cctgtaacgc ctccaagggg tatgatcaca aagcaagcaa agaaataggt cgacggtacc
     2401 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     2461 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2521 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2581 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2641 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2701 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2761 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2821 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2881 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2941 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3001 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3061 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3121 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3181 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3241 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3361 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3541 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3601 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3661 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3721 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3781 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3841 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3901 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3961 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4021 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4081 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4141 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4201 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4261 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4321 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4381 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4441 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4501 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4561 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4621 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4681 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4741 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4801 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4861 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4921 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4981 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5041 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5101 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5161 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5221 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5281 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5341 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5401 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5461 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5521 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5581 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5641 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
//
