LOCUS       sequence-484241-        5331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Ras GTPase, Excitation: 483, Emission: 506.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5331)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 5331)
  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"
COMMENT     Sequence Label: mTagGFP2-Rab5a-7
FEATURES             Location/Qualifiers
     source          1..5331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             547..1260
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     polyA_signal    2059..2180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2096..2115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2150..2169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2187..2642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2324..2345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2536..2555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2669..2773
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2775..3132
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2775..2966
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2798..2818)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2983..3118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3167..3961
                     /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             3167..3961
                     /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(3221..3240)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3831..3850
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4149..4168)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4193..4240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4569..5157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5058..5077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggtc
      541 gccaccatga gcgggggcga ggagctgttc gccggcatcg tgcccgtgct gatcgagctg
      601 gacggcgacg tgcacggcca caagttcagc gtgcgcggcg agggcgaggg cgacgccgac
      661 tacggcaagc tggagatcaa gttcatctgc accaccggca agctgcccgt gccctggccc
      721 accctggtga ccaccctctg ctacggcatc cagtgcttcg cccgctaccc cgagcacatg
      781 aagatgaacg acttcttcaa gagcgccatg cccgagggct acatccagga gcgcaccatc
      841 cagttccagg acgacggcaa gtacaagacc cgcggcgagg tgaagttcga gggcgacacc
      901 ctggtgaacc gcatcgagct gaagggcaag gacttcaagg aggacggcaa catcctgggc
      961 cacaagctgg agtacagctt caacagccac aacgtgtaca tccgccccga caaggccaac
     1021 aacggcctgg aggctaactt caagacccgc cacaacatcg agggcggcgg cgtgcagctg
     1081 gccgaccact accagaccaa cgtgcccctg ggcgacggcc ccgtgctgat ccccatcaac
     1141 cactacctga gcactcagac caagatcagc aaggaccgca acgaggcccg cgaccacatg
     1201 gtgctcctgg agtccttcag cgcctgctgc cacacccacg gcatggacga gctgtacagg
     1261 tccggactca gatctcgagc catggctagt cgaggcgcaa caagacccaa cgggccaaat
     1321 actggaaata aaatatgcca gttcaaacta gtacttctgg gagagtctgc tgttggcaaa
     1381 tcaagcctag tgcttcgttt tgtgaagggc caatttcatg aatttcaaga gagtaccata
     1441 ggggctgctt ttctaaccca aactgtgtgt cttgatgata caacagtaaa gtttgaaata
     1501 tgggatacag ctggtcaaga acgataccat agcttagcac caatgtacta cagaggagca
     1561 caagcagcca tagttgtata tgatatcaca aatgaggagt cctttgccag agccaaaaac
     1621 tgggttaaag aacttcagag gcaagccagt cctaacattg taatagcttt atcaggaaac
     1681 aaggctgatc ttgcaaataa aagagctgtc gatttccagg aagcacagtc ctatgcagat
     1741 gacaacagtt tattattcat ggagacatca gctaaaacat cgatgaatgt aaatgaaata
     1801 ttcatggcaa tagctaaaaa gttgccaaag aacgaaccac agaatccagg agcaaattct
     1861 gccagaggaa gaggagtaga ccttactgaa cccacgcagc caaccaggaa tcagtgttgt
     1921 agtaactaag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     1981 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2041 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2161 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2641 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2761 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2881 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2941 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3061 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3121 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3181 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3241 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3301 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3361 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3421 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3481 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3541 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3601 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3661 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3721 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3781 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3841 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3901 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3961 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4021 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4081 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4141 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4201 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4261 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4321 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4381 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     5281 atagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
