LOCUS       sequence-173185-        5349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Ras GTPase, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5349)
  AUTHORS   Markwardt ML, Kremers GJ, Kraft CA, Ray K, Cranfill PJ, Wilson KA, 
            Day RN, Wachter RM, Davidson MW, Rizzo MA
  TITLE     An improved cerulean fluorescent protein with enhanced brightness 
            and reduced reversible photoswitching.
  JOURNAL   PLoS One. 2011  . 6(3):e17896.
  PUBMED    21479270
REFERENCE   2  (bases 1 to 5349)
  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: "PLoS One. 
            2011  . 6(3):e17896."
COMMENT     Sequence Label: mCerulean3-Rab4a-7
FEATURES             Location/Qualifiers
     source          1..5349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1525..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1562..1581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1616..1635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1653..2108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1790..1811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2002..2021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2135..2239
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2241..2598
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2241..2432
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2264..2284)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2449..2584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2633..3427
                     /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             2633..3427
                     /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(2687..2706)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3297..3316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3615..3634)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3659..3706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4035..4623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4524..4543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4798..5101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5102..5305
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5256..5276
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5344..5349,1..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 gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgagctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaacgccat ccacggcaac gtctatatca ccgccgacaa gcagaagaac
      481 ggcatcaagg ccaacttcgg cctcaactgc aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctcgagccat gtcgcagacg gccatgtccg aaacctacga ttttttgttt
      781 aagttcttgg ttattggaaa tgcaggaact ggcaaatctt gcttacttca tcagtttatt
      841 gaaaaaaaat tcaaagatga ctcaaatcat acaataggag tggaatttgg ttcaaagata
      901 ataaatgttg gtggtaaata tgtaaagtta caaatatggg atacagcagg acaagaacga
      961 ttcaggtccg tgacgagaag ttattaccga ggcgcggccg gggctctcct cgtctatgat
     1021 atcaccagcc gagaaaccta caatgcgctt actaattggt taacagatgc ccgaatgctg
     1081 gcgagccaga acattgtgat catcctttgt ggaaacaaga aggacctgga tgcagatcgt
     1141 gaagttacct tcttagaagc ctccagattt gctcaagaaa atgagctgat gtttttggaa
     1201 acaagtgcgc tcacagggga gaatgtagaa gaggcttttg tacagtgtgc aagaaaaata
     1261 cttaacaaaa tcgaatcagg tgagctggac ccagaaagaa tgggctcagg tattcagtac
     1321 ggagatgctg ccttgagaca gctgaggtca ccgcggcgcg cacaggcccc gaacgctcag
     1381 gagtgtggtt gttagggatc caccggatct agataactga tcataatcag ccataccaca
     1441 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     1501 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     1561 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1621 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     1681 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1741 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1801 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1861 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1921 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1981 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2041 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2101 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2161 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2221 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     2281 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2341 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2401 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2461 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2521 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2581 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     2641 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2701 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2761 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     2821 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2881 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2941 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3001 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3061 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3121 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     3181 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3241 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3301 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3361 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3421 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3481 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3541 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     3601 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3661 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     3721 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3781 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3841 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     3901 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3961 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4021 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4081 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4141 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4201 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4261 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4321 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4381 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4441 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4501 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4561 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4621 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4681 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     4741 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4801 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4861 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4921 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4981 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5041 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     5101 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     5161 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     5221 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     5281 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     5341 gtcgccacc
//
