LOCUS       sequence-321654-        4327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His-tagged, SV40 NLS-tagged tdTomato in the 
            modified pRSETB protein expression backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4327)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 4327)
  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: NLS-tdTomato
FEATURES             Location/Qualifiers
     source          1..4327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             214..234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             235..1665
                     /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"
     primer_bind     complement(1728..1746)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1742..1789
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1886..2341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1973..1992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2183..2204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2367..2471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2472..2540,2541..3332)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2690..2709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3503..4091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3992..4011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4245..4262
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcccgc caccatgact gctccaaaga agaagcgtaa ggtaatggtg
      241 agcaagggcg aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc
      301 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
      361 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
      421 tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat
      481 tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
      541 ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag
      601 gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg
      661 ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc
      721 caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac
      781 atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc
      841 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac
      901 cacctgttcc tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc
      961 tccgaggaca acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
     1021 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1081 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     1141 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     1201 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1261 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
     1321 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
     1381 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
     1441 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc
     1501 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
     1561 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
     1621 cgccaccacc tgttcctgta cggcatggac gagctgtaca agtaagaatt cgaagcttga
     1681 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1741 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1801 aactatatcc ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1861 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1921 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1981 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2041 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2101 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2161 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2221 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2281 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2341 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2461 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2581 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2641 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2761 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2881 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3001 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3061 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3181 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3301 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3361 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3421 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3481 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3541 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3601 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3661 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3721 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3781 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3841 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3901 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3961 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4021 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4081 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4141 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4201 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4261 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     4321 aatgcag
//
