LOCUS       647_pEGFP-RME8_3        5956 bp    DNA     circular SYN 28-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5956)
  AUTHORS   Lobingier Lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
COMMENT     Sequence Label: pEGFP-C1-DNAJC13(351t)-tetramer
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  613 ..  615 / #05fd14
                        2:  616 ..  618 / #05fd14 / 1a
                        3:  619 .. 1329 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1330..1379
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
                     /note="color: #99ccff"
     primer_bind     1375..1432
                     /label=Primer 1
                     /note="color: black; sequence: 
                     TATAGGTACCGGAGGGGGTTCCGGCGGAGGCAGCaacataattagggaaaataaggatc
                     tgg; added: 2021-10-29"
     primer_bind     1375..1394
                     /label=351_F
                     /note="color: black; sequence: TATAGGTACCGGAGGGGGTTCCGG; 
                     added: 2023-04-07"
     CDS             1405..2454
                     /codon_start=1
                     /label=hRME8
                     /note="color: #ff6600"
                     /translation="NIIRENKDLACFYTTKHSWRGKYKRVFSVGTHAITTYNPNTLEVT
                     NQWPYGDICSISPVGKGQGTEFNLTFRKGSGKKSETLKFSTEHRTELLTEALRFRTDFS
                     EGKITGRRYNCYKHHWSDSRKPVILEVTPGGFDQINPATNRVLCSYDYRNIEGFVDLSD
                     YQGGFCILYGGFSRLHLFASEQREEIIKSAIDHAGNYIGISLRIRKEPLEFEQYLNLRF
                     GKYSTDESITSLAEFVVQKISPRHSEPVKRVLALTETCLVERDPATYNIATLKPLGEVF
                     ALVCDSENPQLFTIEFIKGQVRKYSSTERDSLLASLLDGVRASGNRDVCVKMTPTHKGQ
                     RWGLLSMPVD"
     primer_bind     1405..1436
                     /label=FL_N1_GFP_F
                     /note="color: black; sequence: 
                     GCTAGCGCCACCATGGATGaacataattagggaaaataaggatctggcatg; added:
                     2022-11-29"
     primer_bind     2230..2257
                     /label=1
                     /note="color: black; sequence: 
                     CCTTTAGGAGAAGTATTTGCGTTGGTCT; added: 2021-11-29"
     primer_bind     complement(2430..2454)
                     /label=351_R
                     /note="color: black; sequence: 
                     TATAGGATCCCTAatcaacaggcatgctgagtaacccc; added: 2023-04-07"
     CDS             2491..2604
                     /codon_start=1
                     /label=Tetromerization
                     /note="color: #a6acb3"
                     /translation="DYSDLQRVKQELLEEVKKELQKVKEEIIEAFVQELRKR"
     misc_feature    2616..2620
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
                     /note="color: #99ccff"
     misc_feature    2629..2639
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     polyA_signal    2744..2865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2872..3327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3354..3458
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3460..3817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3668..3803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3852..4646
                     /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)"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4878..4925
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5254..5842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 caccctgacc tacggcgtgc agtgcttcag 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 cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 ggagggggtt ccggcggagg cagcaacata attagggaaa ataaggatct ggcatgtttc
     1441 tacacaacaa aacattcatg gagggggaag tataagcgtg tcttttcagt tggaactcat
     1501 gcgattacta catataatcc caatacctta gaagtaacaa atcagtggcc ttatggagac
     1561 atttgcagca tcagccctgt tggaaaagga caaggaacgg agttcaacct cacatttcgt
     1621 aaaggcagtg gaaaaaagtc agaaacttta aaattttcta cagagcacag aacagaactt
     1681 cttacagaag cattgagatt tagaactgat ttttcagagg gaaaaatcac aggaaggaga
     1741 tacaactgct ataagcatca ctggagtgac tcaagaaaac ctgtaatttt ggaagtaact
     1801 ccaggaggct ttgaccaaat taatcctgca accaacagag tactctgttc ctatgactat
     1861 agaaatattg aaggatttgt agatctctca gattatcaag gaggattttg tatactttat
     1921 ggaggattta gtagattgca tttatttgcg tcagagcaaa gagaagagat tattaaaagt
     1981 gcaatagacc atgctggtaa ctacataggt atttcattgc ggatcaggaa agagccttta
     2041 gaattcgagc aatatttgaa tcttcgcttt ggaaaataca gcactgatga atccatcaca
     2101 tctttagcag agtttgtagt ccaaaaaata tcacctagac attcggagcc tgttaaaaga
     2161 gttctagcac ttacagaaac atgtttagta gaacgtgatc cggcaaccta taatattgca
     2221 acattgaagc ctttaggaga agtatttgcg ttggtctgtg actcagaaaa tccacaactt
     2281 tttaccattg aatttataaa agggcaagta cggaaatatt cttcaacaga gagagattcc
     2341 ttattagcaa gtttgctgga tggagtaaga gcctctggta atagagatgt ttgtgtaaaa
     2401 atgacaccaa cccataaagg tcagcgatgg gggttactca gcatgcctgt tgatggcggc
     2461 agcggaggca gtggcgggtc cggggggtca gactattcag atctgcaacg agtgaaacaa
     2521 gagctcctcg aggaggtgaa aaaggagctc cagaaggtaa aggaagaaat aatcgaagcc
     2581 ttcgtccaag aactgagaaa acgctaggcg gccgcgatcc accggatcta gataactgat
     2641 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2701 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2761 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2821 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     2881 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2941 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3001 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3061 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3121 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3181 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3241 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3301 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3421 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3721 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3781 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4021 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4801 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4861 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4921 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4981 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5041 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5101 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5161 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5221 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5281 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5341 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5401 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5461 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5521 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5581 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5641 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5701 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5761 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5821 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5881 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5941 tattaccgcc atgcat
//
