LOCUS       sequence-476906-        8134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 554, Emission: 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8134)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 8134)
  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: tdTomato-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..8134
                     /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..4740
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    4862..4983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4899..4918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4953..4972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4990..5445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5127..5148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5339..5358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5472..5576
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5578..5935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5578..5769
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5601..5621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5786..5921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5970..6764
                     /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             5970..6764
                     /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(6024..6043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6634..6653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6952..6971)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6996..7043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7372..7960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7861..7880
                     /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 tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg
     3361 cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     3421 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     3481 gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc
     3541 gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     3601 atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc
     3661 acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg
     3721 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
     3781 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag
     3841 ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac
     3901 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     3961 tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc
     4021 tccggcaccg cctcctccga ggacaacaac atggccgtca tcaaagagtt catgcgcttc
     4081 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     4141 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     4201 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
     4261 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
     4321 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
     4381 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
     4441 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     4501 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
     4561 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
     4621 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4681 gagcgctccg agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtaa
     4741 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4801 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4861 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4981 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5041 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5101 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5161 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5221 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5281 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5341 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5401 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     5461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5521 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     5581 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     5941 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6001 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6061 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6121 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     6181 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6241 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6301 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6361 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6421 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6481 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6541 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6601 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6661 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6721 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6781 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6841 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6901 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     6961 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     7021 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     7081 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     7141 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     7201 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     7261 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7321 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7381 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7441 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7501 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7561 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7621 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7681 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7741 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7801 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7861 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7921 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7981 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8041 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     8101 ggtcattagt tcatagccca tatatggagt tccg
//
