LOCUS       sequence-306363-        6540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 554, Emission: 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6540)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 6540)
  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-Actin-C-18
FEATURES             Location/Qualifiers
     source          1..6540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      613..622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             619..2046
                     /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    3406..3527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3497..3516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3534..3989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3883..3902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4016..4120
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4145..4165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4189..4399
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4211..4407
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4258..4393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4442..5236
                     /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             4442..5236
                     /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(4496..4515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5106..5125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5424..5443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5468..5515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5844..6432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6333..6352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
      661 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      781 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
      841 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
      961 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     1021 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     1081 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     1141 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     1201 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1261 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc
     1321 agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat caaagagttc
     1381 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     1441 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     1501 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg
     1561 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc
     1621 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc
     1681 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     1741 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     1801 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
     1861 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     1921 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     1981 gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg
     2041 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
     2101 ggcccgggat ctggcagcgg tggaggcagc gcatccggcg gaagcggaag cgatgatgat
     2161 atcgccgcgc tcgtcgtcga caacggctcc ggcatgtgca aggccggctt cgcgggcgac
     2221 gatgcccccc gggccgtctt cccctccatc gtggggcgcc ccaggcacca gggcgtgatg
     2281 gtgggcatgg gtcagaagga ttcctatgtg ggcgacgagg cccagagcaa gagaggcatc
     2341 ctcaccctga agtaccccat cgagcacggc atcgtcacca actgggacga catggagaaa
     2401 atctggcacc acaccttcta caatgagctg cgtgtggctc ccgaggagca ccccgtgctg
     2461 ctgaccgagg cccccctgaa ccccaaggcc aaccgcgaga agatgaccca gatcatgttt
     2521 gagaccttca acaccccagc catgtacgtt gctatccagg ctgtgctatc cctgtacgcc
     2581 tctggccgta ccactggcat cgtgatggac tccggtgacg gggtcaccca cactgtgccc
     2641 atctacgagg ggtatgccct cccccatgcc atcctgcgtc tggacctggc tggccgggac
     2701 ctgactgact acctcatgaa gatcctcacc gagcgcggct acagcttcac caccacggcc
     2761 gagcgggaaa tcgtgcgtga cattaaggag aagctgtgct acgtcgccct ggacttcgag
     2821 caagagatgg ccacggctgc ttccagctcc tccctggaga agagctacga gctgcctgac
     2881 ggccaggtca tcaccattgg caatgagcgg ttccgctgcc ctgaggcact cttccagcct
     2941 tccttcctgg gcatggagtc ctgtggcatc cacgaaacta ccttcaactc catcatgaag
     3001 tgtgacgtgg acatccgcaa agacctgtac gccaacacag tgctgtctgg cggcaccacc
     3061 atgtaccctg gcattgccga caggatgcag aaggagatca ctgccctggc acccagcaca
     3121 atgaagatca agatcattgc tcctcctgag cgcaagtact ccgtgtggat cggcggctcc
     3181 atcctggcct cgctgtccac cttccagcag atgtggatca gcaagcagga gtatgacgag
     3241 tccggcccct ccatcgtcca ccgcaaatgc ttctagggat ccaccggatc tagataactg
     3301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3541 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3601 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3661 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3721 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3781 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3841 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3901 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3961 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4081 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4141 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4201 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4261 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4321 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4381 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4441 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4501 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4561 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4621 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4681 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4741 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4801 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4861 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4921 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4981 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5041 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5101 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5161 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5221 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5281 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5341 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5401 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5461 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5521 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5581 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5641 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
//
