LOCUS       sequence-402562-        4756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pPP2AA3 promoter with GG0 and GG1 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4756)
  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 4756)
  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-I2 pPP2AA3-mutated GG0-GG1
FEATURES             Location/Qualifiers
     source          1..4756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             594..605
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1357..1373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1357..1373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1370..1392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1381..1397
                     /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(1405..1411,1412..1429,1430..1435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1450..1471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1588..1605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1759..2347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1839..1858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2485..2859)
                     /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      2856..2865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3065..3859)
                     /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(3065..3859)
                     /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(3176..3195)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3786..3805
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(4208..4510)
                     /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(4310..4329)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     4633..4655
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4647..4664
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4648..4664
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4671..4690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4671..4689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactctgtgg tctcaggagg aggaagaaga agagagttac tgagaagctt cctttatacg
       61 agagagatat taaacgaagt cgtatttgtt ggatatatgg gcctttatgt attatacata
      121 agcccattta gttcttatct ttgacaggcc caattatatg gacacagctt gaagatggtt
      181 tagctgctgc gaaagacgag ctccggtctg atttctcgtt cttctgattg atagatcgct
      241 cggaacttgg aaagcagcgt aatcggtaaa ttctcgatca tttcttcaat ttcgaaacat
      301 tttccaattg tatattgaga atctcaaatc gaattattgt ttagatcttc tctctttatt
      361 agtctaagaa tggtaatagt ttctctctag attgggacgg cacaaagttt cagttctcca
      421 ttctaatcat tttttagagg ctaatgggtc acctttgaat tatctcagag gcggaattga
      481 gtttaaagtt tgtaacttta tcgtcttttg tattgatggt gtctatggca aaggtgatta
      541 actgcaaagg tatgcattgt gttttattct gactttgtgt acatttgtgt ttgattgaag
      601 gtagggagtg gcaaaggtga ttaactgcaa aggatttgag ttttggtgag gaatgtgaga
      661 ccatcaacgt agtcagaata aaacacaatg catacctttg cagttaatca cctttgccat
      721 agacaccatc aatacaaaag acgataaagt tacaaacttt aaactcaatt ccgcctctga
      781 gataattcaa aggtgaccca ttagcctcta aaaaatgatt agaatggaga actgaaactt
      841 tgtgccgtcc caatctagag agaaactatt accattctta gactaataaa gagagaagat
      901 ctaaacaata attcgatttg agattctcaa tatacaattg gaaaatgttt cgaaattgaa
      961 gaaatgatcg agaatttacc gattacgctg ctttccaagt tccgagcgat ctatcaatca
     1021 gaagaacgag aaatcagacc ggagctcgtc tttcgcagca gctaaaccat cttcaagctg
     1081 tgtccatata attgggcctg tcaaagataa gaactaaatg ggcttatgta taatacataa
     1141 aggcccatat atccaacaaa tacgacttcg tttaatatct ctctcgtata aaggaagctt
     1201 ctcagtaact ctcttcttct tcctcctcct gagaccacag agtgcctgaa ttccagcaca
     1261 ctggcggccg ttactagtgg atccgagctc ggtaccaagc ttgatgcata gcttgagtat
     1321 tctaacgcgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1381 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1621 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2461 ttaaaaatga agttttagca cgtgtcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2521 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2581 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2641 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2701 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2761 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2821 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tcacgtgcta
     2881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
     3001 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     3061 taattcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     3121 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     3181 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     3241 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     3301 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     3361 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     3421 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     3481 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     3541 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     3601 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     3661 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     3721 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     3781 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     3841 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gagcttgatc
     3901 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     3961 caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg
     4021 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     4081 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     4141 gcggactggc tttctacgtg aaaaggatct aggtgaagat cctttttgat aatctcatgc
     4201 ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt
     4261 ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg
     4321 tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt
     4381 tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccc ggcgtgtcaa
     4441 taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa
     4501 ccttaaactg ccgtacgtat aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4561 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     4621 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     4681 cactataggg cgaattgggc cctctagatg catgctcgag cggccgccag tgtgatggat
     4741 atctgcagaa ttcagg
//
