LOCUS       sequence-221561-        6411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  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 6411)
  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-Paxillin-22
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2217..2269
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2277..2286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2283..3014
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3139..3260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3176..3195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3230..3249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3267..3722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3616..3635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3749..3853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3855..4212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3855..4046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3878..3898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4063..4198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4247..5041
                     /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             4247..5041
                     /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(4301..4320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4911..4930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5229..5248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5273..5320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5649..6237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6138..6157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggacgacct cgatgcctta ctggcagacc tggaatccac cacctcgcat atctctaagc
      601 gaccggtgtt tctaacagag gaaacgcctt actcctaccc aactggaaac catacgtacc
      661 aggagattgc tgtgccaccg ccggtgcccc cgccgccttc cagtgaggcc ctaaatggca
      721 ctgtgattga ccccttagac cagtggcaac ccagcgtgtc cagatacggc caccagcaac
      781 ctcagtccca gtctccgata tacagctcta gtgccaaaag ctccagtgcc tctgttccta
      841 gagacgggct cagctcccct tctccccgtg ccagtgagga ggaacacgtg tacagcttcc
      901 caaacaagca gaagtctgca gaaccatctc ccacaatgac cagcacctcc ctgggcagca
      961 acctctcgga actggacagg cttctcctgg aactgaatgc tgttcaacat aatcccccca
     1021 gtggcttctc agcagatgaa gtcagcagaa gcccatcact gcccaatgtg accggacctc
     1081 actatgtcat cccagagagc agcagctctg caggagggaa ggctgcaccc cctacaaaag
     1141 aaaaaccaaa gcggaatggt gggcgtggga ttgaagatgt gcgtcccagt gtggagagcc
     1201 tgctggatga gctggagagc tctgtgccaa gcccagtccc tgcaatcact gtgagccaag
     1261 gggaggtgag cagcccccag cgtgtcaacg ccagtcagca gcagacccgt atctctgctt
     1321 cttcagctac acgagaactg gatgagctga tggcgtccct ctctgacttt aagttcatgg
     1381 cacaggggaa agccggcggc agcagctccc caccatccac caccccaaag cctggcagtc
     1441 agctggacac catgctggga agtctgcagt ctgacctgaa caaactgggg gtagcgacgg
     1501 tcgccaaggg tgtgtgcgga gcctgcaaga agccgattgc tgggcaggta gttacagcca
     1561 tggggaagac ctggcaccct gagcacttcg tctgcaccca ctgccaggag gagattggat
     1621 cacggaactt ctttgagcgg gatgggcagc cctactgtga gaaggactat cacaacctct
     1681 tctctcctcg ctgctactac tgcaatgggc caatccttga taaagtggtg acggctttgg
     1741 acaggacgtg gcaccctgaa cactttttct gtgcccagtg tggagttttc tttggacctg
     1801 aaggatttca tgagaaggat ggcaaagcct attgccgcaa ggactacttt gacatgtttg
     1861 ctcccaagtg tggaggctgt gcccgggcta tcctggaaaa ttacatttct gccttaaaca
     1921 ccctgtggca ccctgagtgc tttgtctgtc gggaatgttt cacaccgttc atcaacggca
     1981 gcttcttcga gcacgacggg cagccctact gcgaggtgca ttaccacgag cgccgcggct
     2041 cgctgtgctc tggctgccag aagcccatca cgggacgctg catcactgcc atgggcaaga
     2101 aatttcaccc cgaacacttt gtctgtgcct tctgcctcaa gcagctcaac aaaggaacct
     2161 tcaaagaaca gaacgacaag ccctactgtc agaactgctt tctcaaactc ttctgtctcg
     2221 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2281 ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg
     2341 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     2401 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     2461 acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca
     2521 tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     2581 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     2641 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     2701 aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag
     2761 gcagaaccga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga
     2821 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
     2881 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
     2941 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt aatggcatgg
     3001 acgagctgta caagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3061 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3121 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3241 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3841 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4261 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4321 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4381 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4441 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4501 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4561 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4621 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4681 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4741 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4801 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4861 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4921 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4981 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5041 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5101 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5161 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5221 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5281 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5341 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5401 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5461 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5521 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5581 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5641 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5701 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5761 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5821 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5881 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5941 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6001 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6061 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6301 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     6361 atagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
