LOCUS       sequence-176713-        5340 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Ras GTPase, Excitation: 505, Emission: 515.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 5340)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: Clover-Rab5a-7
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..717
                     /codon_start=1
                     /product="monomeric derivative of the bright green-yellow 
                     fluorescent protein Clover"
                     /label=mClover2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1516..1637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1553..1572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1607..1626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1644..2099)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1781..1802)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1993..2012
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2126..2230
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2232..2589
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2232..2423
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2255..2275)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2440..2575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2624..3418
                     /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             2624..3418
                     /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(2678..2697)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3288..3307
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3606..3625)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3650..3697
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4026..4614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4515..4534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4789..5092
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5093..5296
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5247..5267
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5335..5340,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct
      301 ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaacttcaa cagccacaac gtctatatca cggccgacaa gcagaagaac
      481 ggcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagcc atcagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctcgagccat ggctagtcga ggcgcaacaa gacccaacgg gccaaatact
      781 ggaaataaaa tatgccagtt caaactagta cttctgggag agtctgctgt tggcaaatca
      841 agcctagtgc ttcgttttgt gaagggccaa tttcatgaat ttcaagagag taccataggg
      901 gctgcttttc taacccaaac tgtgtgtctt gatgatacaa cagtaaagtt tgaaatatgg
      961 gatacagctg gtcaagaacg ataccatagc ttagcaccaa tgtactacag aggagcacaa
     1021 gcagccatag ttgtatatga tatcacaaat gaggagtcct ttgccagagc caaaaactgg
     1081 gttaaagaac ttcagaggca agccagtcct aacattgtaa tagctttatc aggaaacaag
     1141 gctgatcttg caaataaaag agctgtcgat ttccaggaag cacagtccta tgcagatgac
     1201 aacagtttat tattcatgga gacatcagct aaaacatcga tgaatgtaaa tgaaatattc
     1261 atggcaatag ctaaaaagtt gccaaagaac gaaccacaga atccaggagc aaattctgcc
     1321 agaggaagag gagtagacct tactgaaccc acgcagccaa ccaggaatca gtgttgtagt
     1381 aactaaggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     1441 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1501 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1561 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1621 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     1681 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1741 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1801 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1861 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1921 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1981 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2041 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2101 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2161 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2221 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2281 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2341 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2401 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2461 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2521 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2581 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2641 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2701 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2761 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     2821 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2881 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     2941 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3001 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3061 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3121 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3181 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3241 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3301 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3361 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3421 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3481 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3541 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     3601 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3661 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3721 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3781 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3841 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     3901 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3961 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4021 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4081 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4141 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4201 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4261 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4321 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4381 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4441 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4501 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4561 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4621 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4681 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     4741 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4801 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4861 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4921 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4981 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5041 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5101 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5161 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5221 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     5281 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
//
