LOCUS       sequence-468018-        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane/Vesicles, Excitation: 462, Emission: 492.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  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 6299)
  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-C-Src-7
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             635..676
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     regulatory      2249..2258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2255..2965
                     /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    3087..3208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3178..3197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3215..3670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3352..3373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3564..3583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3697..3801
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3803..4160
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3803..3994
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3826..3846)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4011..4146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4195..4989
                     /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             4195..4989
                     /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(4249..4268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4859..4878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5177..5196)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5221..5268
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5597..6185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6086..6105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgggg agcagcaaga gcaagcccaa
      661 ggaccccagc cagcgccggc gcagcctgga gccacccgac agcacccacc acgggggatt
      721 cccagcctcg cagaccccca acaagacagc agcccccgac acgcaccgca cccccagccg
      781 ctcctttggg accgtggcca ccgagcccaa gctcttcggg ggcttcaaca cttctgacac
      841 cgtcacgtcg ccgcagcgtg ccggggcact ggctggcggc gtcaccactt tcgtggctct
      901 ctatgactac gagtcccgga ctgaaacgga cttgtccttc aagaaaggag aacgcctgca
      961 gattgtcaac aacacggaag gtgactggtg gctggctcat tccctcacta caggacagac
     1021 gggctacatc cccagtaact atgtcgcgcc ctcagactcc atccaggctg aagagtggta
     1081 ctttgggaag atcactcgtc gggagtccga gcggctgctg ctcaaccccg aaaacccccg
     1141 gggaaccttc ttggtccggg agagcgagac gacaaaaggt gcctattgcc tctccgtttc
     1201 tgactttgac aacgccaagg ggctcaatgt gaagcactac aagatccgca agctggacag
     1261 cggcggcttc tacatcacct cacgcacaca gttcagcagc ctgcagcagc tggtggccta
     1321 ctactccaaa catgctgatg gcttgtgcca ccgcctgacc aacgtctgcc ccacgtccaa
     1381 gccccagacc cagggactcg ccaaggacgc gtgggaaatc ccccgggagt cgctgcggct
     1441 ggaggtgaag ctggggcagg gctgctttgg agaggtctgg atggggacct ggaacggcac
     1501 caccagagtg gccataaaga ctctgaagcc cggcaccatg tccccggagg ccttcctgca
     1561 ggaagcccaa gtgatgaaga agctccggca tgagaagctg gttcagctgt acgcagtggt
     1621 gtcggaagag cccatctaca tcgtcactga gtacatgagc aaggggagcc tcctggattt
     1681 cctgaaggga gagatgggca agtacctgcg gctgccacag ctcgtcgata tggctgctca
     1741 gattgcatcc ggcatggcct atgtggagag gatgaactac gtgcaccgag acctgcgggc
     1801 ggccaacatc ctggtggggg agaacctggt gtgcaaggtg gctgactttg ggctggcacg
     1861 cctcatcgag gacaacgagt acacagcacg gcaaggtgcc aagttcccca tcaagtggac
     1921 agcccccgag gcagccctct atggccggtt caccatcaag tcggatgtct ggtccttcgg
     1981 catcctgctg actgagctga ccaccaaggg ccgggtgcca tacccaggga tggtcaacag
     2041 ggaggtgctg gaccaggtgg agaggggcta ccgcatgccc tgcccgcccg agtgccccga
     2101 gtcgctgcat gacctcatgt gccagtgctg gcggaaggac cctgaggagc ggcccacttt
     2161 tgagtacctg caggccttcc tggaggacta cttcacctcg acagagcccc agtaccagcc
     2221 tggagagaac ctacgggatc caccggtcgc caccatggtg agcaagggcg aggagaccac
     2281 aatgggcgta atcaagcccg acatgaagat caagctgaag atggagggca acgtgaatgg
     2341 ccacgccttc gtgatcgagg gcgagggcga gggcaagccc tacgacggca ccaacaccat
     2401 caacctggag gtgaaggagg gagcccccct gcccttctcc tacgacattc tgaccaccgc
     2461 gttcgcctac ggcaacaggg ccttcaccaa gtaccccgac gacatcccca actacttcaa
     2521 gcagtccttc cccgagggct actcttggga gcgcaccatg accttcgagg acaagggcat
     2581 cgtgaaggtg aagtccgaca tctccatgga ggaggactcc ttcatctacg agatacacct
     2641 caagggcgag aacttccccc ccaacggccc cgtgatgcag aagaagacca ccggctggga
     2701 cgcctccacc gagaggatgt acgtgcgcga cggcgtgctg aagggcgacg tcaagcacaa
     2761 gctgctgctg gagggcggcg gccaccaccg cgttgacttc aagaccatct acagggccaa
     2821 gaaggcggtg aagctgcccg actatcactt tgtggaccac cgcatcgaga tcctgaacca
     2881 cgacaaggac tacaacaagg tgaccgttta cgagagcgcc gtggcccgca actccaccga
     2941 cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca
     3001 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3061 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3181 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3241 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3661 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3721 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3781 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3841 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3961 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4021 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4081 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4141 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5101 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5161 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5221 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5281 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5341 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5401 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
