LOCUS       sequence-226428-        7438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7438)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 7438)
  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-T2-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..7438
                     /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      3304..3313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3310..4041
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    4166..4287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4203..4222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4257..4276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4294..4749)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4431..4452)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4643..4662
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4776..4880
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4882..5239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4882..5073
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4905..4925)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5090..5225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5274..6068
                     /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             5274..6068
                     /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(5328..5347)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5938..5957
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6256..6275)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6300..6347
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6676..7264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7165..7184
                     /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 ccggactcag atctcgagct caagcttcgc accatcatgg accattatga ttctcagcaa
      601 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      661 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      721 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      781 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      841 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      901 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
      961 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1021 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1081 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1141 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1201 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1261 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1321 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1381 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1441 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1501 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1561 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1621 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1681 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1741 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1801 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1861 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1921 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     1981 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2041 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2101 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2161 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2221 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2281 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2341 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2401 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2461 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2521 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2581 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2641 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2701 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2761 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2821 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2881 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     2941 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3001 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3061 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3121 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3181 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3241 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3301 gtcgccacca tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg
     3361 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
     3421 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
     3481 gccttcgaca tcctggctac cagcttcatg tacggcagca gaaccttcat caaccacacc
     3541 cagggcatcc ccgatttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     3601 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     3661 tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg
     3721 cagaagaaaa cactcggctg ggaggccaac accgagaccc tgtaccccgc tgacggcggc
     3781 ctggaaggca gaaccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     3841 ttcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
     3901 tatgtggacc acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
     3961 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat
     4021 ggcatggacg agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac
     4081 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4141 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4261 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     4321 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4381 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4441 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4501 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4561 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4621 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4681 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4741 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4801 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4861 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4921 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4981 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5041 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5101 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5161 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5221 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5281 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5341 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5401 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5461 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5521 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5581 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5641 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5701 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5761 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5821 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5881 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5941 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6001 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6061 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6121 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6181 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6241 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6301 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6361 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6421 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6481 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     7381 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
