LOCUS       646_pEGFP-RME8_3        5947 bp    DNA     circular SYN 28-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5947)
  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)-dimer
FEATURES             Location/Qualifiers
     source          1..5947
                     /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..2595
                     /codon_start=1
                     /label=dimerization
                     /note="color: #a6acb3"
                     /translation="EIAALKKEIAALKQENAALKQEIAALKKEIAALKQ"
     misc_feature    2607..2611
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
                     /note="color: #99ccff"
     misc_feature    2620..2630
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     polyA_signal    2735..2856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2863..3318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3345..3449
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3451..3808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3659..3794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3843..4637
                     /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    4869..4916
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5245..5833
                     /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 gaaatagccg ccttgaagaa agaaatcgca
     2521 gcgcttaaac aagagaatgc agcgcttaaa caggagatcg ccgccctcaa gaaggagata
     2581 gccgcactca agcagtaggc ggccgcgatc caccggatct agataactga tcataatcag
     2641 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2701 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2761 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2821 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     2881 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2941 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3001 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3061 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3121 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3181 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3241 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3301 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3421 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3481 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3601 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3661 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3721 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3781 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3841 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3901 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3961 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4021 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4081 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4141 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4201 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4261 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4321 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4381 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4441 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4501 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4561 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4621 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4681 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4741 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4801 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4861 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4921 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4981 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5041 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5101 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5161 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5221 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5281 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5341 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5401 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5461 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5521 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5581 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5641 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5701 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5761 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5821 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5881 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5941 catgcat
//
