LOCUS       sequence-386291-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pUBQ10 promoter with GG0 and GG1 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5576)
  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 5576)
  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-I26 pUBQ10 GG0-GG1
FEATURES             Location/Qualifiers
     source          1..5576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2131..2150)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2132..2150)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2157..2174)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2157..2173)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2166..2188)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2311..2613
                     /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     2492..2511
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2962..3756
                     /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             2962..3756
                     /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(3016..3035)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3626..3645
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3956..3965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3962..4336
                     /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      4474..5062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4963..4982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5216..5233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5350..5371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5386..5391,5392..5409,5410..5416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5424..5440
                     /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     5429..5451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5448..5464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5448..5464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg tctcaggagc tcaacaacaa actttccatt cacccattta tggatacaca
       61 gtctagctca acagagcttt taacccaaat tggtacaata gaatacaact ttagatcata
      121 attctcaaaa gaaagagatt ccttagctat tctatctgcc actccatttc cttctcggct
      181 tgtatgcaca agcataaaat cctcaaactt gctaagtaga tactttatgt cttggataat
      241 tggattgaga cttgacaagc ataactttca tgtaaccaaa gacacaagtt gctgagaatc
      301 cacctcaaaa atgatcttcc tataattgaa tcgggataat gacagcacag cccatctaag
      361 agcctccact tctacttcca gcacgcttct tacttttacc acagctcttg cacctaacca
      421 taacaccttc cctgtatgat cgcgaagcac ccaccctaag ccacatttta atccttctgt
      481 tggccatgcc ccatcaaagt tgcacttaac ccaagattgt ggtggagctt cccatgtttc
      541 tcgtctgtcc cgacggtgtt gtggttggtg ctttccttac attctgagcc tctttccttc
      601 taatccactc atctgcatct tcttgtgtcc ttactaatac ctcattggtt ccaaattccc
      661 tccctttaag caccagctcg tttctgttct tccacagcct cccaagtatc caagggacta
      721 aagcctccac attcttcaga tcaggatatt cttgtttaag atgttgaact ctatggaggt
      781 ttgtatgaac tgatgatcta ggaccggata agttcccttc ttcatagcga acttattcaa
      841 agaatgtttt gtgtatcatt cttgttacat tgttattaat gaaaaaatat tattggtcat
      901 tggactgaac acgagtgtta aatatggacc aggccccaaa taagatccat tgatatatga
      961 attaaataac aagaataaat cgagtcacca aaccacttgc cttttttaac gagacttgtt
     1021 caccaacttg atacaaaagt cattatccta tgcaaatcaa taatcataca aaaatatcca
     1081 ataacactaa aaaattaaaa gaaatggata atttcacaat atgttatacg ataaagaagt
     1141 tacttttcca agaaattcac tgattttata agcccacttg cattagataa atggcaaaaa
     1201 aaaacaaaaa ggaaaagaaa taaagcacga agaattctag aaaatacgaa atacgcttca
     1261 atgcagtggg acccacggtt caattattgc caattttcag ctccaccgta tatttaaaaa
     1321 ataaaacgat aatgctaaaa aaatataaat cgtaacgatc gttaaatctc aacggctgga
     1381 tcttatgacg accgttagaa attgtggttg tcgacgagtc agtaataaac ggcgtcaaag
     1441 tggttgcagc cggcacacac gagtcgtgtt tatcaactca aagcacaaat acttttcctc
     1501 aacctaaaaa taaggcaatt agccaaaaac aactttgcgt gtaaacaacg ctcaatacac
     1561 gtgtcatttt attattagct attgcttcac cgccttagct ttctcgtgac ctagtcgtcc
     1621 tcgtcttttc ttcttcttct tctataaaac aatacccaaa gagctcttct tcttcacaat
     1681 tcagatttca atttctcaaa atcttaaaaa ctttctctca attctctcta ccgtgatcaa
     1741 ggtaaatttc tgtgttcctt attctctcaa aatcttcgat tttgttttcg ttcgatccca
     1801 atttcgtata tgttctttgg tttagattct gttaatctta gatcgaagac gattttctgg
     1861 gtttgatcgt tagatatcat cttaattctc gattagggtt tcatagatat catccgattt
     1921 gttcaaataa tttgagtttt gtcgaataat tactcttcga tttgtgattt ctatctagat
     1981 ctggtgttag tttctagttt gtgcgatcga atttgtcgat taatctgagt ttttctgatt
     2041 aacagaatgt gagaccatca acgtcctgaa ttctgcagat atccatcaca ctggcggccg
     2101 ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta caattcactg
     2161 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2221 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2281 tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta taaaagagag
     2341 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc ggggcgacgg
     2401 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac
     2461 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     2521 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     2581 aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt atcaaaaagg
     2641 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     2701 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     2761 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     2821 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     2881 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag
     2941 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3001 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3061 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3121 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3181 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3241 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3301 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3361 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3421 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3481 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3541 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3601 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3661 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3721 atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg
     3781 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     3841 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3901 tccgctcatg agacaataac cctgataaat gcttcaataa tagcacgtga ggagggccac
     3961 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     4021 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     4081 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     4141 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     4201 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     4261 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     4321 cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga tctaggtgaa
     4381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5161 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5221 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5281 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5341 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5401 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     5461 tgaccatgat tacgccaagc tatttaggtg acgcgttaga atactcaagc tatgcatcaa
     5521 gcttggtacc gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcagg
//
