LOCUS       sequence-386439-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pUBQ10 promoter between BpiI sites for target construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  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 5579)
  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-I38 BpiI-pUBQ10-BpiI
FEATURES             Location/Qualifiers
     source          1..5579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2134..2153)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2135..2153)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2160..2177)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2160..2176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2169..2191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2314..2616
                     /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     2495..2514
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2965..3759
                     /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             2965..3759
                     /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(3019..3038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3629..3648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3959..3968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3965..4339
                     /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      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4966..4985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5219..5236
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5353..5374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5389..5394,5395..5412,5413..5419)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5427..5443
                     /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     5432..5454
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5451..5467
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5451..5467
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg aagacaagga gctcaacaac aaactttcca ttcacccatt tatggataca
       61 cagtctagct caacagagct tttaacccaa attggtacaa tagaatacaa ctttagatca
      121 taattctcaa aagaaagaga ttccttagct attctatctg ccactccatt tccttctcgg
      181 cttgtatgca caagcataaa atcctcaaac ttgctaagta gatactttat gtcttggata
      241 attggattga gacttgacaa gcataacttt catgtaacca aagacacaag ttgctgagaa
      301 tccacctcaa aaatgatctt cctataattg aatcgggata atgacagcac agcccatcta
      361 agagcctcca cttctacttc cagcacgctt cttactttta ccacagctct tgcacctaac
      421 cataacacct tccctgtatg atcgcgaagc acccacccta agccacattt taatccttct
      481 gttggccatg ccccatcaaa gttgcactta acccaagatt gtggtggagc ttcccatgtt
      541 tctcgtctgt cccgacggtg ttgtggttgg tgctttcctt acattctgag cctctttcct
      601 tctaatccac tcatctgcat cttcttgtgt ccttactaat acctcattgg ttccaaattc
      661 cctcccttta agcaccagct cgtttctgtt cttccacagc ctcccaagta tccaagggac
      721 taaagcctcc acattcttca gatcaggata ttcttgttta agatgttgaa ctctatggag
      781 gtttgtatga actgatgatc taggaccgga taagttccct tcttcatagc gaacttattc
      841 aaagaatgtt ttgtgtatca ttcttgttac attgttatta atgaaaaaat attattggtc
      901 attggactga acacgagtgt taaatatgga ccaggcccca aataagatcc attgatatat
      961 gaattaaata acaagaataa atcgagtcac caaaccactt gcctttttta acgagacttg
     1021 ttcaccaact tgatacaaaa gtcattatcc tatgcaaatc aataatcata caaaaatatc
     1081 caataacact aaaaaattaa aagaaatgga taatttcaca atatgttata cgataaagaa
     1141 gttacttttc caagaaattc actgatttta taagcccact tgcattagat aaatggcaaa
     1201 aaaaaacaaa aaggaaaaga aataaagcac gaagaattct agaaaatacg aaatacgctt
     1261 caatgcagtg ggacccacgg ttcaattatt gccaattttc agctccaccg tatatttaaa
     1321 aaataaaacg ataatgctaa aaaaatataa atcgtaacga tcgttaaatc tcaacggctg
     1381 gatcttatga cgaccgttag aaattgtggt tgtcgacgag tcagtaataa acggcgtcaa
     1441 agtggttgca gccggcacac acgagtcgtg tttatcaact caaagcacaa atacttttcc
     1501 tcaacctaaa aataaggcaa ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac
     1561 acgtgtcatt ttattattag ctattgcttc accgccttag ctttctcgtg acctagtcgt
     1621 cctcgtcttt tcttcttctt cttctataaa acaataccca aagagctctt cttcttcaca
     1681 attcagattt caatttctca aaatcttaaa aactttctct caattctctc taccgtgatc
     1741 aaggtaaatt tctgtgttcc ttattctctc aaaatcttcg attttgtttt cgttcgatcc
     1801 caatttcgta tatgttcttt ggtttagatt ctgttaatct tagatcgaag acgattttct
     1861 gggtttgatc gttagatatc atcttaattc tcgattaggg tttcatagat atcatccgat
     1921 ttgttcaaat aatttgagtt ttgtcgaata attactcttc gatttgtgat ttctatctag
     1981 atctggtgtt agtttctagt ttgtgcgatc gaatttgtcg attaatctga gtttttctga
     2041 ttaacagtac tttgtcttca tcaacgtcct gaattctgca gatatccatc acactggcgg
     2101 ccgctcgagc atgcatctag agggcccaat tcgccctata gtgagtcgta ttacaattca
     2161 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2221 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2281 ccttcccaac agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga
     2341 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga
     2401 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     2461 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     2521 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     2581 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa
     2641 aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc
     2701 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     2761 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     2821 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     2881 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa
     2941 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3001 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3061 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3121 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3181 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3241 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3301 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3361 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3421 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3481 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3541 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3601 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3661 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3721 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc
     3781 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac
     3841 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3901 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatagcacg tgaggagggc
     3961 caccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     4021 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     4081 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     4141 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     4201 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     4261 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     4321 ggccgaggag caggactgac acgtgctaaa acttcatttt taatttaaaa ggatctaggt
     4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5101 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5161 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5221 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5281 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5341 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     5401 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5461 ctatgaccat gattacgcca agctatttag gtgacgcgtt agaatactca agctatgcat
     5521 caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg gaattcagg
//
