LOCUS       sequence-267742-        5367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Clathrin Vesicles, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 5367)
  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"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mTagRFP-T-Clathrin-15
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      653..662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             659..1390
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2201..2322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2238..2257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2292..2311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2329..2784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2466..2487)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2678..2697
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2811..2915
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2917..3274
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2917..3108
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2940..2960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3125..3260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3309..4103
                     /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             3309..4103
                     /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(3363..3382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3973..3992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4291..4310)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4335..4382
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4711..5299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5200..5219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
      661 ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg
      721 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
      781 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
      841 cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc
      901 cgatttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
      961 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     1021 caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac
     1081 actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag
     1141 aaccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac
     1201 atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca
     1261 cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc
     1321 tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatg gcatggacga
     1381 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacatggc
     1441 tgatgacttt ggcttcttct cgtcgtcgga gagtggtgcc ccggaggcgg cggaggagga
     1501 cccggcggcc gccttcctgg cccagcagga gagcgagatt gcaggcatag agaacgacga
     1561 gggcttcggg gcacctgccg gcagccatgc ggcccccgca cagccgggcc ccacgagtgg
     1621 ggctggttct gaggacatgg ggaccacagt caatggagat gtgtttcagg aggccaacgg
     1681 tcctgctgat ggctacgcag ccattgccca ggctgacagg ctgacccagg agcctgagag
     1741 catccgcaag tggcgagagg agcagaggaa acggctgcaa gagctggatg ctgcatctaa
     1801 ggtcacggaa caggaatggc gggagaaggc caagaaggac ctggaggagt ggaaccagcg
     1861 ccagagtgaa caagtagaga agaacaagat caacaaccgg gcatccgagg aggctttcgt
     1921 gaaggaatcc aaggaggaga ccccaggcac agagtgggag aaggtggccc agctatgtga
     1981 cttcaacccc aagagcagca agcagtgcaa agatgtgtcc cgcctgcgct cggtgctcat
     2041 gtccctgaag cagacgccac tgtcccgcta gggatccacc ggatctagat aactgatcat
     2101 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2161 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2281 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2341 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2401 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2461 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2521 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2581 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2641 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2701 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2761 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2821 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2881 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2941 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3121 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3181 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3241 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4141 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4261 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4321 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4381 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4441 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4501 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5341 ctggcctttt gctcacatgt tctttcc
//
