LOCUS       sequence-115699-        5443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 554, Emission: 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5443)
  AUTHORS   Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien 
            RY
  TITLE     Improved monomeric red, orange and yellow fluorescent proteins 
            derived from Discosoma sp. red fluorescent protein.
  JOURNAL   Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.
  PUBMED    15558047
REFERENCE   2  (bases 1 to 5443)
  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: "Nat 
            Biotechnol."; date: "2004-12"; volume: "22(12)"; pages: "1567-72. 
            Epub 2004 Nov 21"
COMMENT     Sequence Label: tdTomato-N1
FEATURES             Location/Qualifiers
     source          1..5443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    598..678
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      680..689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             686..2116
                     /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    2238..2359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2275..2294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2329..2348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2366..2821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2503..2524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2715..2734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2848..2952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2954..3311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2954..3145
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2977..2997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3162..3297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3346..4140
                     /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             3346..4140
                     /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(3400..3419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4010..4029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4328..4347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4372..4419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4748..5336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5237..5256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgaca
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
      661 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt
      721 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
      781 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      841 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
      901 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
      961 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     1021 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     1081 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     1141 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     1201 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     1261 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     1321 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     1381 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa
     1441 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     1501 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1561 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     1621 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     1681 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     1741 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     1801 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     1861 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     1921 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     1981 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     2041 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga
     2101 cgagctgtac aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2161 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2221 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2341 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2401 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2461 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2521 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2581 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2641 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2701 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2761 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2941 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3181 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3241 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3301 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3361 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3421 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3481 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3541 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3601 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3661 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3721 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3781 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3841 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3901 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3961 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4021 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4081 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4141 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4201 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4261 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4321 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4381 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4441 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4501 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4561 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4621 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4681 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4741 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4801 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4861 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4921 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4981 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5041 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5101 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5161 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5221 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5281 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5341 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5401 ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
