LOCUS       sequence-198671-        5821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin/Focal Adhesions, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5821)
  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: mTagRFP-T-VASP-5
FEATURES             Location/Qualifiers
     source          1..5821
                     /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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1284
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2549..2670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2586..2605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2640..2659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2677..3132)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2814..2835)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3026..3045
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3159..3263
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3265..3622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3265..3456
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3288..3308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3473..3608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3657..4451
                     /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             3657..4451
                     /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(3711..3730)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4321..4340
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4639..4658)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4683..4730
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5059..5647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5548..5567
                     /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 gctagcgcta
      541 ccggtcgcca ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
      601 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
      661 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      721 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac
      781 acccagggca tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga
      841 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      901 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
      961 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc
     1021 ggcctggaag gcagaaccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1081 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1141 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1201 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     1261 aatggcatgg acgagctgta caagtccgga ctcagatcta tgagcgagac ggtcatctgt
     1321 tccagccggg ctactgtgat gctttatgat gacagcaaca agcgatggct ccctgctggc
     1381 actggtcccc aggccttcag ccgcgtccag atataccaca accccactgc taactccttc
     1441 cgagttgttg gccgcaagat gcagccggat cagcaggtgg ttatcaactg tgccatcatt
     1501 cggggtgtca agtacaatca ggccactccc atcttccatc agtggcgaga tgcccgccag
     1561 gtctggggcc tcaacttcgg cagcaaggag gacgccatac agtttgcaac aggcatggcc
     1621 aacgccctag aggccttgga aggaggtggg cctcccccag ccccagcacc ccctgcctgg
     1681 tctgcccaga atggtccctc cccagaggag ctggaacaac agaaaaggca gccggagcat
     1741 atggagcgcc gggtctccaa tgcaggaggc ccacctgctc ccccagctgg gggccctcct
     1801 ccacctccag gacctccccc tcctccaggt ccccccccac ccccaggtct gccctcctca
     1861 ggggtatctg gggcaggtca tggagcaggg gcagccccac cccctgcacc cccactccct
     1921 acaacacagg gccccaatag tgggggttcc ggggccccag gcctggctgc tgccattgct
     1981 ggagccaaac tcaggaaagt gagcaagcag gaggaggcct ctggggggcc cctggccccc
     2041 aaagctgaga acagtcgaag cactggtggg gggcttatgg aagagatgaa cgccatgctg
     2101 gcccggagaa gaaaagccac acaggttggg gagaagcccc ccaaagacga gtcagccagt
     2161 gaggagtcag aggcccgact ccctgcccag agtgaacctg tgagaagacc ctgggagaag
     2221 aacagcacaa ccttgccaag gatgaagtcg tcctcttctg tgactacctc cgaggcccac
     2281 ccctcgacgc cctgctccag tgatgactcc gacttggaga gggtgaagca ggagcttctg
     2341 gaagaggtgc ggaaggagct acagaaaatg aaagaggaaa tcatcgaagt ctttgtccag
     2401 gagctgagga agcggggttc tccttgagcg gccgcgactc tagatcataa tcagccatac
     2461 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2521 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2581 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2641 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     2701 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2761 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2821 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2881 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2941 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3001 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3061 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3121 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3181 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3241 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3301 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3361 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3421 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3481 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3541 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3601 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3661 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3721 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3781 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3841 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3901 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3961 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4021 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4081 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4141 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4201 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4261 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4321 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4381 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4441 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4501 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4561 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4621 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4681 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4741 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4801 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4861 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     4921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5701 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5761 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5821 g
//
