LOCUS       sequence-241202-        5340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Ras GTPase, Excitation: 462, Emission: 492.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  AUTHORS   Ai HW, Olenych SG, Wong P, Davidson MW, Campbell RE
  TITLE     Hue-shifted monomeric variants of Clavularia cyan fluorescent 
            protein: identification of the molecular determinants of color and 
            applications in fluorescence imaging.
  JOURNAL   BMC Biol. 2008 Mar 6;6:13. doi: 10.1186/1741-7007-6-13.
  PUBMED    18325109
REFERENCE   2  (bases 1 to 5340)
  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"; journalName: "BMC Biol.";
            date: "2008-03-6"; volume: "6:13. doi"; pages: " 
            10.1186/1741-7007-6-13"
COMMENT     Sequence Label: mTFP1-Rab4a-7
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        151..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     609..629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             703..1410
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    2218..2339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2255..2274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2309..2328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2346..2801)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2483..2504)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2695..2714
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2828..2932
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2934..3291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2934..3125
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2957..2977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3142..3277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3326..4120
                     /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             3326..4120
                     /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(3380..3399)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3990..4009
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4308..4327)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4352..4399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4728..5316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5217..5236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
       61 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
      121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      661 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      721 gagaccacaa tgggcgtaat caagcccgac atgaagatca agctgaagat ggagggcaac
      781 gtgaatggcc acgccttcgt gatcgagggc gagggcgagg gcaagcccta cgacggcacc
      841 aacaccatca acctggaggt gaaggaggga gcccccctgc ccttctccta cgacattctg
      901 accaccgcgt tcgcctacgg caacagggcc ttcaccaagt accccgacga catccccaac
      961 tacttcaagc agtccttccc cgagggctac tcttgggagc gcaccatgac cttcgaggac
     1021 aagggcatcg tgaaggtgaa gtccgacatc tccatggagg aggactcctt catctacgag
     1081 atacacctca agggcgagaa cttccccccc aacggccccg tgatgcagaa gaagaccacc
     1141 ggctgggacg cctccaccga gaggatgtac gtgcgcgacg gcgtgctgaa gggcgacgtc
     1201 aagcacaagc tgctgctgga gggcggcggc caccaccgcg ttgacttcaa gaccatctac
     1261 agggccaaga aggcggtgaa gctgcccgac tatcactttg tggaccaccg catcgagatc
     1321 ctgaaccacg acaaggacta caacaaggtg accgtttacg agagcgccgt ggcccgcaac
     1381 tccaccgacg gcatggacga gctgtacaag tccggactca gatctcgagc catgtcgcag
     1441 acggccatgt ccgaaaccta cgattttttg tttaagttct tggttattgg aaatgcagga
     1501 actggcaaat cttgcttact tcatcagttt attgaaaaaa aattcaaaga tgactcaaat
     1561 catacaatag gagtggaatt tggttcaaag ataataaatg ttggtggtaa atatgtaaag
     1621 ttacaaatat gggatacagc aggacaagaa cgattcaggt ccgtgacgag aagttattac
     1681 cgaggcgcgg ccggggctct cctcgtctat gatatcacca gccgagaaac ctacaatgcg
     1741 cttactaatt ggttaacaga tgcccgaatg ctggcgagcc agaacattgt gatcatcctt
     1801 tgtggaaaca agaaggacct ggatgcagat cgtgaagtta ccttcttaga agcctccaga
     1861 tttgctcaag aaaatgagct gatgtttttg gaaacaagtg cgctcacagg ggagaatgta
     1921 gaagaggctt ttgtacagtg tgcaagaaaa atacttaaca aaatcgaatc aggtgagctg
     1981 gacccagaaa gaatgggctc aggtattcag tacggagatg ctgccttgag acagctgagg
     2041 tcaccgcggc gcgcacaggc cccgaacgct caggagtgtg gttgttaggg atccaccgga
     2101 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2161 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2221 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2281 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2341 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2401 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2461 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2521 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2581 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2641 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2701 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2761 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     2941 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3121 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3181 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3241 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3301 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3361 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3421 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3481 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3541 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3601 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3661 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3721 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3781 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3841 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3901 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3961 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4021 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4081 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4141 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4201 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4261 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4321 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4381 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4441 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4501 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4561 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4621 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4681 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4741 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4801 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4861 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4921 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4981 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5041 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5101 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5161 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5221 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5281 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
//
