LOCUS       sequence-345228-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SYS1-mEGFP can be transfected in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  AUTHORS   Ishida M, Bonifacino JS
  TITLE     ARFRP1 functions upstream of ARL1 and ARL5 to coordinate recruitment
            of distinct tethering factors to the trans-Golgi network.
  JOURNAL   J Cell Biol. 2019 Nov 4;218(11):3681-3696. doi: 
            10.1083/jcb.201905097. Epub 2019 Oct 1.
  PUBMED    31575603
REFERENCE   2  (bases 1 to 5184)
  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"; doi: 
            "10.1083/jcb.201905097"; journalName: "J Cell Biol"; date: 
            "2019-11-4- 4"; volume: "218"; issue: "11"; pages: "3681-3696"
COMMENT     Sequence Label: pmEGFP-N1 SYS1
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..245
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     202..222
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(247..351)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      378..833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(465..484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     675..696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(851..870)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     905..924
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1084..1797,1798..1800,1801..1803))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1129..1150)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1478..1497
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1737..1758
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1800..1809
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2289..2298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2365..2568)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2394..2414)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2569..2872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3047..3635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3127..3146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3964..4011
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4036..4055
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4243..5037)
                     /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             complement(4243..5037)
                     /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(4354..4373)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4964..4983
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5072..5184,1..245))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5086..5184,1..37)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
       61 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      121 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      181 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
      241 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
      301 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
      361 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
      721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcctt
      841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      961 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1021 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
     1081 gctttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     1141 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact gggtgctcag
     1201 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1261 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1321 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1381 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1441 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1501 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1561 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     1621 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     1681 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     1741 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     1801 catggtggcg accggtggat cccggacatt ggatttaggg gctgagttga ggggtatctc
     1861 cttgagctcc gtccgcatgc acaggtactc cccgatgaca gccatgagtg caatgcacac
     1921 ggcttggacc agccaccagg tcagcgccga ggggaaacgg gagctgtaga accagcagcc
     1981 caggaggtga aagaaatgga cagtgacagt gaaatccaga cactgctttc ctcgccggat
     2041 gaagtacagc aagcccaggg cacaggtgag ggcgttgagg atgaaggaca tcatggagag
     2101 ccggcctgga ggggtggaaa agcccaggat ctcggcgtcg aacatctggt ccagcgaggg
     2161 gctgcttcgc actagcccgt ccaccagcgc cagccacagg cccagcgagc cgtaatacac
     2221 ggtctgcatg aggacgatct gcgacaggat cagcagcggg tcccacacgt agctgcggaa
     2281 ctgacccgcc atggtggcga attcgaagct tgagctcgag atctgagtcc ggtagcgcta
     2341 gcggatctga cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct
     2401 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     2461 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     2521 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     2581 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     2641 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     2701 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     2761 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     2821 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     2881 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac taatgcatgg
     2941 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3001 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3061 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3121 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3181 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3241 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3301 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3361 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3421 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3481 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3541 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3601 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3661 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3721 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3781 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     3841 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     3901 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     3961 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     4021 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     4081 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     4141 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     4201 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     4261 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4321 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4381 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4441 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4501 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     4561 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     4621 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     4681 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4741 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4801 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     4861 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4921 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4981 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     5041 aaacgatcct catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc
     5101 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5161 aaaaaattag tcagccatgg ggcg
//
