LOCUS       sequence-386303-        3683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 DST (destabilization RNA tag: SAUR tag) with GG4 GG5 golden 
            gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3683)
  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 3683)
  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-I14 DST GG4-GG5
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     145..162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    279..300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(315..320,321..338,339..345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    353..369
                     /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     358..380
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     377..393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     377..393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(743..762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(744..762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(769..786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(769..785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(778..800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             923..1225
                     /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     1104..1123
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1574..2368
                     /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             1574..2368
                     /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(1628..1647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2238..2257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2568..2577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2574..2948
                     /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      3086..3674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3575..3594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
       61 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      121 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      181 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      241 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      301 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      361 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct atttaggtga
      421 cgcgttagaa tactcaagct atgcatcaag cttggtaccg agctcggatc cactagtaac
      481 ggccgccagt gtgctggaat tcaggacgtt gatggtctca taccaacatg caaatacgca
      541 ttgatcaatt acgatctatg tattgtatat tcctctcatt gaaacaattt acaattatct
      601 ccaggggaaa ttagctaaca atctaaaaaa aatatggaaa tgttgtagta tagtactaag
      661 ctgagacctc cgatcccctg aattctgcag atatccatca cactggcggc cgctcgagca
      721 tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
      781 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      841 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      901 gttgcgcagc ctatacgtac ggcagtttaa ggtttacacc tataaaagag agagccgtta
      961 tcgtctgttt gtggatgtac agagtgatat tattgacacg ccggggcgac ggatggtgat
     1021 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
     1081 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
     1141 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     1201 taacctgatg ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac
     1261 ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     1321 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     1381 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     1441 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     1501 tttcttgccg ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg
     1561 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     1621 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     1681 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     1741 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     1801 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     1861 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     1921 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     1981 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2041 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2101 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2161 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2221 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2281 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2341 ctatcgcctt cttgacgagt tcttctgaat tattaacgct tacaatttcc tgatgcggta
     2401 ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa
     2461 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2521 tgagacaata accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca
     2581 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     2641 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     2701 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     2761 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     2821 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     2881 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     2941 aggactgaca cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3001 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3061 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3661 ggagcctatg gaaaaacgcc agc
//
