LOCUS       sequence-386977-        3797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 Empty Tp901 target: attP_Tp901-BpiI-attB_Tp901 with golden 
            gate linker in pBLUNT plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3797)
  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 3797)
  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-T7 Empty target Tp901
FEATURES             Location/Qualifiers
     source          1..3797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(358..374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(358..374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(371..393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    382..398
                     /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(406..412,413..430,431..436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    451..472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(589..606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(760..1348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(840..859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1486..1860)
                     /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      1857..1866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2066..2860)
                     /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(2066..2860)
                     /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(2177..2196)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2787..2806
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3209..3511)
                     /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(3311..3330)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3634..3656
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3648..3665
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3649..3665
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3672..3691
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3672..3690
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactctgtgg tctcactatg cgagttttta tttcgtttat ttcaattaag gtaactaaaa
       61 aactcctttg gagttgtctt cttgtagcac tgtaagattt atccacgaag gtcagcaact
      121 tgaagacaat actgcaaaaa aagcaaaaag catttacctt gattgagatg ttaattgtgt
      181 tggcatacct tgagacctcc gatcccctga attccagcac actggcggcc gttactagtg
      241 gatcccctga attccagcac actggcggcc gttactagtg gatccgagct cggtaccaag
      301 cttgatgcat agcttgagta ttctaacgcg tcacctaaat agcttggcgt aatcatggtc
      361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1441 cttcacctag atccttttaa attaaaaatg aagttttagc acgtgtcagt cctgctcctc
     1501 ggccacgaag tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg
     1561 ctgctcgccg atctcggtca tggccggccc ggaggcgtcc cggaagttcg tggacacgac
     1621 ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc cacacccagg ccagggtgtt
     1681 gtccggcacc acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac
     1741 accggcgaag tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg tccagaactc
     1801 gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg gcactggtca acttggccat
     1861 ggtggccctc ctcacgtgct attattgaag catttatcag ggttattgtc tcatgagcgg
     1921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1981 aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2041 tcaggaaatt gtaagcgtta ataattcaga agaactcgtc aagaaggcga tagaaggcga
     2101 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     2161 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     2221 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     2281 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     2341 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     2401 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     2461 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     2521 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     2581 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     2641 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     2701 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     2761 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     2821 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     2881 tctcttgatc agagcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     2941 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     3001 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     3061 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     3121 tccggggtca gcaccgtttc tgcggactgg ctttctacgt gaaaaggatc taggtgaaga
     3181 tcctttttga taatctcatg cctgacattt atattcccca gaacatcagg ttaatggcgt
     3241 ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg
     3301 gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg
     3361 ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg gggatcacca
     3421 tccgtcgccc cggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc
     3481 tctctctttt ataggtgtaa accttaaact gccgtacgta taggctgcgc aactgttggg
     3541 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     3601 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3661 ccagtgaatt gtaatacgac tcactatagg gcgaattggg ccctctagat gcatgctcga
     3721 gcggccgcca gtgtgatgga tatctgcaga attcaggcga gcggccgcca gtgtgatgga
     3781 tatctgcaga attcagg
//
