LOCUS       sequence-176185-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4743)
  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: 
            "unpublished"
COMMENT     Sequence Label: mTagRFP-T-Lifeact-7
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             190..984
                     /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             190..984
                     /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(244..263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     854..873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1172..1191)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1216..1263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1592..2180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2081..2100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2355..2658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2659..2862
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2813..2833
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2891..2900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2897..2947
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      2963..2972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2969..3700
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    3825..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3862..3881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3916..3935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3953..4408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4090..4111)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4302..4321
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4435..4539
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4541..4743,1..155)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4541..4732
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4564..4584)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
       61 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      121 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
      181 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
      241 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
      301 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
      361 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
      421 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
      481 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
      541 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
      601 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
      661 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
      721 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
      781 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
      841 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
      901 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
      961 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     1021 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     1081 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     1141 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     1201 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     1261 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     1321 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     1381 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     1441 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1501 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1561 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1621 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1681 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1741 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1801 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1861 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1921 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1981 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2041 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2101 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2161 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2221 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2281 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     2341 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2401 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2461 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2521 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2581 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     2641 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     2701 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2761 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2821 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     2881 atccgctagc gccaccatgg gcgtggccga cttgatcaag aagttcgagt ccatctccaa
     2941 ggaggagggg gatccaccgg tcgccaccat ggtgtctaag ggcgaagagc tgattaagga
     3001 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     3061 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     3121 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag
     3181 aaccttcatc aaccacaccc agggcatccc cgatttcttt aagcagtcct tccctgaggg
     3241 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     3301 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     3361 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagatgct
     3421 gtaccccgct gacggcggcc tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg
     3481 gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct
     3541 caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa
     3601 agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     3661 actggggcac aaacttaatg gcatggacga gctgtacaag taagcggccg cgactctaga
     3721 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3781 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3841 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3901 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3961 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4021 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4081 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4141 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4201 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4261 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4321 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4381 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4561 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4621 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4681 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4741 ctc
//
