LOCUS       sequence-386438-        4941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 tRBSC-mScarlet for integrase target construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  AUTHORS   Guiziou S, Maranas CJ, Chu JC, Nemhauser JL
  TITLE     An integrase toolbox to record gene-expression during plant 
            development.
  JOURNAL   Nat Commun. 2023 Apr 3;14(1):1844. doi: 10.1038/s41467-023-37607-5.
  PUBMED    37012288
REFERENCE   2  (bases 1 to 4941)
  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 
            Commun."; date: "2023-04-3"; volume: "14(1):1844. doi"; pages: " 
            10.1038/s41467-023-37607-5"
COMMENT     Sequence Label: L0-T4 tRBSC_mScarlet
FEATURES             Location/Qualifiers
     source          1..4941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(23..664)
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     CDS             complement(673..1365)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(948..967)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1217..1235
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1381..1401)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1496..1515)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1497..1515)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1522..1539)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1522..1538)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1531..1553)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1676..1978
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1857..1876
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2327..3121
                     /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             2327..3121
                     /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(2381..2400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2991..3010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3321..3330
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3327..3701
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3839..4427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4328..4347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4581..4598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4715..4736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4751..4756,4757..4774,4775..4781)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4789..4805
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4794..4816
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4813..4829
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4813..4829
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg tctcaggagc gattgatgca tgttgtcaat caattggcaa gtcataaaat
       61 gcattaaaaa atattttcat actcaactac aaatccatga gtataactat aattataaag
      121 caatgattag aatctgacaa ggattctgga aaattacata aaggaaagtt cataaatgtc
      181 taaaacacaa gaggacatac ttgtattcag taacatttgc agcttttcta ggtctgaaaa
      241 tatatttgtt gcctagtgaa taagcataat ggtacaacta caagtgtttt actcctcata
      301 ttaacttcgg tcattagagg ccacgatttg acacattttt actcaaaaca aaatgtttgc
      361 atatctctta tattttcaaa ttcaacacac aacaaataag agaaaaaaca aataatatta
      421 atttgagaat gaacaaaagg accatatcat tcattaactc ttctccatcc atttccattt
      481 cacagttcga tagcgaaaac cgaataaaaa acacagtaaa ttacaagcac aacaaatggt
      541 acaagaaaaa catgtttccc aatgccataa tactcgaact cagtaggatt ctggtgtgtg
      601 cgcaatgaaa ctgatgcatt gaacttgacg aacgttgtcg aaaccgatga tacgaacgaa
      661 agctcaagct tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggagcg
      721 ttcgtactgt tccaccacgg tgtagtcctc gttgtgggag gtgatgtcca acttgcggtc
      781 gacgttgtag gcgccgggca tctgcacggg cttcttggcc ttgtaggtgg tcttgaagtc
      841 cgccaggtag cggccgccgt ccttcaggcg cagggccatc ttaatgtcgc ccttcagcac
      901 gccgtcctcg gggtacaacc gctcggtgga cgcttcccag cccattgtct tcttctgcat
      961 tacggggccg tcaggaggga agttggtgcc gcggagcttc accttgtaga tcagggtgcc
     1021 gtcctccagg gaggtgtcct gggtcacggt cacggcgccg ccgtcctcga agttcatcac
     1081 gcgctcccac ttgaagccct cggggaagga ctgcttatag tagtcgggga tgtcggcggg
     1141 gtgcttggtg aaggccctgg agccgtacat gaactgaggg gacaggatgt cccaggagaa
     1201 gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg
     1261 gccctcgccc tcgccctcga tctcgaactc gtggccgttc atggagccct ccatgtgcac
     1321 cttgaaccgc atgaactcct tgatcactgc ctcgcccttg ctcacaccac taccacttcc
     1381 gactttcctc ttcttctttg gcattgttta ctattgagac catcaacgtc ctgaattctg
     1441 cagatatcca tcacactggc ggccgctcga gcatgcatct agagggccca attcgcccta
     1501 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1561 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1621 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg tacggcagtt
     1681 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1741 tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1801 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     1861 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     1921 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     1981 gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt agaaagccag
     2041 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     2101 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     2161 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     2221 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     2281 caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2341 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2401 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2461 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2521 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2581 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2641 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2701 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2761 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2821 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     2881 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2941 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3001 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3061 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3121 aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3181 ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3241 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3301 aataatagca cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt ccggtgctca
     3361 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     3421 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     3481 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     3541 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     3601 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     3661 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta aaacttcatt
     3721 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3961 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4501 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4561 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4621 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4681 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     4741 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     4801 aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt aggtgacgcg
     4861 ttagaatact caagctatgc atcaagcttg gtaccgagct cggatccact agtaacggcc
     4921 gccagtgtgc tggaattcag g
//
