LOCUS       sequence-388004-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pLBD16 promoter with GG0 and GG1 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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-I32 pLBD16 GG0-GG1
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    290..311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(326..331,332..349,350..356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    364..380
                     /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     369..391
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     388..404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     388..404
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1623..1642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1624..1642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1649..1666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1649..1665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1658..1680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1803..2105
                     /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     1984..2003
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2454..3248
                     /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             2454..3248
                     /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(2508..2527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3118..3137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3448..3457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3454..3828
                     /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      join(3966..4552,1..2)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4455..4474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
       61 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      181 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      241 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      301 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      361 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      421 tatttaggtg acgcgttaga atactcaagc tatgcatcaa gcttggtacc gagctcggat
      481 ccactagtaa cggccgccag tgtgctggaa ttcaggcact ctgtggtctc aggagtgcca
      541 ttattgattg tctccttaga atccaacatg tgaccctcca ttgtctacca ttcacaccct
      601 ttggctcact cgataaattt ggacattaat tgcattcatt ttatttgtaa gaaccagaat
      661 ctcagtagat gttatttgtg ttgagtaaac tgttgttatt tatagactag gctattcttc
      721 gtatacactt tgatactgat ttttagttca tcatctatct accgactaaa tatagtgttt
      781 ctatgtggct tctcctcaaa tgtcaaattt atttctttaa tgttgtttga gtttcaaact
      841 tgcaaagtgc aaaccaattt tggttgttat actaaaacca cacatataat taaagacacc
      901 cactatagca taaaattcgt cgtgtaagca attttactat agtctaaact ccataaaagg
      961 acgtcgaaaa atattgtgat attggaatta tagtgtcagt ttgtttttct ttgggcaaat
     1021 ggattgtagt ttcattatat attacaattt ataaacgaaa cgataagtat ggctcatact
     1081 ttagtgatgg agaattaaag tattttatat tcagagatga cacaatatgg tagaccaaaa
     1141 ataactaatt aaaatagtat catattagat atttatatgt gattgtaaat ttgatcattg
     1201 agtgaacatt ttaaattagt atgcccaaga aagaaaagaa aaaaaaaaaa ctctttctct
     1261 ctcatgataa tgttcctttt ataaaatcca cccaaagaca ccaaacactt ctccatcatc
     1321 atttcatttc ttgttctttt tccaccatca ccatttccta gttcgctcgt gcacatctcc
     1381 gacgatccca attctaaaat catcacttct tccaatcttt tcaacatctc ttcaaaatat
     1441 ccagtcaaca aaaaaaaaag tacttaaaac aaggaaaccc cttctttctt tttgtacata
     1501 taacataaaa atcatcactt ttttgttcgt ttcgccgaat gtgagaccac gaagtgcctg
     1561 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggcccaatt
     1621 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     1681 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1741 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac
     1801 ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     1861 agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc
     1921 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     1981 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     2041 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     2101 tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga
     2161 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     2221 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     2281 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     2341 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     2401 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg
     2461 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2521 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2581 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     2641 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2701 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2761 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2821 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2881 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2941 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     3001 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3061 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3121 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3181 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3241 tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3301 gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatagcacgt gaggagggcc accatggcca agttgaccag tgccgttccg
     3481 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     3541 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     3601 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     3661 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     3721 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     3781 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctaaaa
     3841 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4081 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4141 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4201 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4261 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4321 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4381 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4441 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4501 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg ga
//
