LOCUS       sequence-457779-        5316 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 5316)
  TITLE     Davidson Cyan Fluorescent Proteins
REFERENCE   2  (bases 1 to 5316)
  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: mCerulean-Rab7a-7
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(10..367)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      24..159
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        176..367
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     324..344
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(369..473)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      500..955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(587..606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     797..818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    962..1083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(973..992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1027..1046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1858..2568,2569..2571,2572..2574))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1900..1921)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2249..2268
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2508..2529
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2571..2580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2619..2822)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2648..2668)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2823..3126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3301..3889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3381..3400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4218..4265
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4290..4309
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4497..5291)
                     /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             complement(4497..5291)
                     /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(4608..4627)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5218..5237
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
       61 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
      121 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
      181 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      241 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      301 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
      361 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      481 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      901 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcg
      961 ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1141 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gttatctaga
     1201 tccggtggat cctcagcaac tgcagctttc tgccgaggcc ttggcccggt cattcttgtc
     1261 cagtttgata ggttcaggaa attcgttgta cagctccacc tccgtttcct gcttaagtgc
     1321 attccgtgca atcgtctgga acgcctgctc cacgttgatg gcctccttgg cactggtctc
     1381 aaagtaggga atgttgtttt tgctgtagca ccaggcctgt gcccgctttg tggccacttg
     1441 tctgttttcg aggtcaatct tgtttcccaa cacaacaaat gggaagtttt caggatctcg
     1501 gggactggcc tggatgagaa actcatctct ccagctatct agggttttga atgtgttggg
     1561 ggcagtcaca tcaaatacca gaacgcagca gtctgcacct ctgtagaagg ccacaccgag
     1621 agactggaac cgttcctgtc ctgctgtgtc ccatatctgc attgtgacta gcctgtcatc
     1681 caccatcacc tccttggtca gaaagtcagc tcctattgtg gctttgtact gattgctgaa
     1741 tttcttattc acatactggt tcatgagtga tgtcttcccg actccagaat ctcccaggat
     1801 gataaccttc agcaacactt tcttcctaga ggtcatggct cgagatctga gtccggactt
     1861 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     1921 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt
     1981 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     2041 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt
     2101 ctgcttgtcg gcggtgatat agacgttgtc gctgatggcg ttgtactcca gcttgtgccc
     2161 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     2221 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     2281 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     2341 gtggtcgggg tagcgggcga agcactgcac gccccaggtc agggtggtca cgagggtggg
     2401 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     2461 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     2521 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     2581 gaccggtagc gctagcggat ctgacggttc actaaaccag ctctgcttat atagacctcc
     2641 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     2701 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2761 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     2821 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     2881 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2941 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     3001 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     3061 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     3121 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     3181 taactaatgc atggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3721 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4021 atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt
     4081 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac
     4141 gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga
     4201 ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat
     4261 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     4321 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4381 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     4441 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     4501 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4561 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4621 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     4681 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     4741 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc
     4801 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     4861 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     4921 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     4981 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5041 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5101 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5161 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5221 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5281 tgttcaatca tgcgaaacga tcctcatcct gtctct
//
