LOCUS       sequence-386342-        4050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 tUBQ1 terminator with GG5 GG6 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4050)
  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 4050)
  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-I16 tUBQ1 GG5-GG6
FEATURES             Location/Qualifiers
     source          1..4050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(651..667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(651..667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(664..686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    675..691
                     /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)."
     promoter        complement(join(699..705,706..723,724..729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    744..765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(882..899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1053..1641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1133..1152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1779..2153)
                     /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"
     regulatory      2150..2159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2359..3153)
                     /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             complement(2359..3153)
                     /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(2470..2489)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3080..3099
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3502..3804)
                     /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     complement(3604..3623)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3927..3949
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3941..3958
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3942..3958
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3965..3984
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3965..3983
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactctgtgg tctcaggtaa gagactctta tcaagaatcc catctcttgc ttgctttttt
       61 ttgttgtctt ccctttgata gggtttgttt ttcttgtttc agtgactttc tatgttaaaa
      121 gataatgtca gtaaaaggat ttggttttct attattctga atcgattacg gaagattctt
      181 gcttaattcc aatctataca agtatcgtga aataatgacc gtttatgtga ttaggagacg
      241 tgtttcatta ataaaatata agatcaatac attgttagta gtgataaact atgtacaaat
      301 tgtattgatt gtaaaagaaa cacaataggt tccttttttc tacaatatat tgtgacagac
      361 tctctgtttt aacgaatgaa ttaaatttgt caacgaaaaa attaaagaat gatgtgatca
      421 ctttgcacct tggttcatct tcagaggctt ctgcatgttc catgcattga ggctctctac
      481 ggtcacaggt tgtgagccgt tgttaaagac aaacaaatgc gcttgagacc tccgatcccc
      541 tgaattccag cacactggcg gccgttacta gtggatccga gctcggtacc aagcttgatg
      601 catagcttga gtattctaac gcgtcaccta aatagcttgg cgtaatcatg gtcatagctg
      661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      781 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1741 tagatccttt taaattaaaa atgaagtttt agcacgtgtc agtcctgctc ctcggccacg
     1801 aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg
     1861 ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac
     1921 cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc
     1981 accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg
     2041 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct
     2101 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtggcc
     2161 ctcctcacgt gctattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2221 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2281 ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     2341 attgtaagcg ttaataattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     2401 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     2461 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     2521 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     2581 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     2641 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     2701 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     2761 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     2821 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     2881 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     2941 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     3001 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3061 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3121 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     3181 atcagagctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     3241 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     3301 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     3361 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     3421 tcagcaccgt ttctgcggac tggctttcta cgtgaaaagg atctaggtga agatcctttt
     3481 tgataatctc atgcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat
     3541 gtcattttcg cggtggctga gatcagccac ttcttccccg ataacggaga ccggcacact
     3601 ggccatatcg gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag
     3661 ttcacgggag actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg
     3721 ccccggcgtg tcaataatat cactctgtac atccacaaac agacgataac ggctctctct
     3781 tttataggtg taaaccttaa actgccgtac gtataggctg cgcaactgtt gggaagggcg
     3841 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3901 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3961 attgtaatac gactcactat agggcgaatt gggccctcta gatgcatgct cgagcggccg
     4021 ccagtgtgat ggatatctgc agaattcagg
//
