LOCUS       sequence-386967-        5368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 PhiC31 integrase with GG2 GG4 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  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 5368)
  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-I10 PhiC31 GG2-GG4
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..1834
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="DTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDGG
                     RFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIPI
                     VSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQRELG
                     GYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWREI
                     KTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRILR
                     DPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQAW
                     LDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPGQ
                     HEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERAN
                     LVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAEA
                     PKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPIE
                     KRASITWAKPPTDDDEDDAQDGTEDVAA"
     primer_bind     complement(1969..1985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1969..1985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1982..2004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1993..2009
                     /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(2017..2023,2024..2041,2042..2047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2062..2083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2200..2217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2371..2959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2451..2470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3097..3471)
                     /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      3468..3477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3677..4471)
                     /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(3677..4471)
                     /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(3788..3807)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4398..4417
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(4820..5122)
                     /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(4922..4941)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     5245..5267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5259..5276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5260..5276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5283..5302
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5283..5301
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactctgtgg tctcaagccg acacgtatgc cggcgcttat gatagacaat caagagaaag
       61 agagaactct tctgccgcct ctccagctac acagagatct gccaacgagg ataaagctgc
      121 tgatttacaa agagaggtag agagagatgg gggtaggttt agatttgtgg gtcatttctc
      181 tgaggctcct ggtacatcag cctttggtac tgctgaacgg cctgaatttg aaagaatatt
      241 aaatgagtgt cgggccggtc gactaaatat gataatagtt tatgatgtat ctaggttcag
      301 tagactcaaa gttatggatg caattccgat cgtgagcgag ctacttgcac tcggtgttac
      361 tatcgtaagt acacaagaag gtgtattcag gcagggtaat gtgatggacc ttatacacct
      421 tattatgcga ctcgacgcta gccataaaga gtcatcactg aagagtgcta aaatcttgga
      481 caccaagaac ctgcaaaggg agcttggagg ctatgtagga ggaaaagctc cgtatggttt
      541 cgaactcgtg agtgagacaa aagagattac cagaaatggt aggatggtga acgtggttat
      601 taataagcta gctcatagca caacccctct cacagggcct tttgagttcg aacctgacgt
      661 tattagatgg tggtggagag agatcaaaac tcacaaacat cttccattta agccaggtag
      721 ccaggctgct atccaccctg gatctatcac tgggctttgt aaaaggatgg atgctgacgc
      781 agtgccaact cgaggggaaa caatcggtaa gaagactgca tcttccgctt gggatcccgc
      841 tacagttatg agaatccttc gggatccgcg tattgctgga ttcgccgcag aagtcattta
      901 taagaaaaaa cctgatggta ctcctactac aaaaatcgaa ggttatagga tccaaaggga
      961 tcctattact ttgcgtccag ttgaattaga ctgtggaccg atcatcgaac ctgccgagtg
     1021 gtacgagttg caggcctggc ttgatggtag agggcgtgga aagggtttat ctcggggaca
     1081 ggcaattttg tcagcaatgg ataagcttta ttgtgagtgt ggggctgtta tgacatcaaa
     1141 acgtggagaa gagtcaatca aggattctta caggtgcaga cgtcgtaaag tggtggaccc
     1201 ttctgctcca ggacagcatg agggtacatg caacgtttct atggctgctc ttgataaatt
     1261 tgtggccgaa cgtattttca acaagattag acacgctgaa ggggatgagg aaactttagc
     1321 tcttctttgg gaggctgccc ggcgttttgg taagctaaca gaggcaccag aaaaaagtgg
     1381 cgaaagagct aatttggtag ccgaaagagc agacgccttg aatgcattgg aagaacttta
     1441 cgaagatagg gctgccggag catatgatgg accagttggt cggaagcatt tcaggaagca
     1501 acaagctgca ctaactttga gacagcaagg cgcagaggag cgactcgcag agcttgaggc
     1561 cgccgaggcc ccaaagcttc cactcgatca atggtttcct gaggatgccg acgctgatcc
     1621 tacaggtcca aagagctggt ggggaagagc tagcgtagac gataagagag tgtttgtggg
     1681 tctatttgtt gacaagattg tagttactaa gtccactacg ggtcgaggac agggcactcc
     1741 catcgaaaag agagccagta ttacctgggc aaagcctcca accgatgatg atgaagacga
     1801 tgcacaagat ggtaccgagg atgtggcagc ataataagct ttgagacctc atcacgcctg
     1861 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca
     1921 tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt
     1981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2101 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3001 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3061 gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa
     3121 gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc
     3181 gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca
     3241 ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac
     3301 cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa
     3361 gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc
     3421 ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct
     3481 cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3541 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3601 acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     3661 tgtaagcgtt aataattcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3721 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3781 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3841 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     3901 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     3961 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4021 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4081 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4141 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4201 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4261 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     4321 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4381 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4441 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4501 cagagcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     4561 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     4621 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     4681 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     4741 agcaccgttt ctgcggactg gctttctacg tgaaaaggat ctaggtgaag atcctttttg
     4801 ataatctcat gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     4861 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
     4921 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     4981 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     5041 ccggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     5101 tataggtgta aaccttaaac tgccgtacgt ataggctgcg caactgttgg gaagggcgat
     5161 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     5221 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     5281 tgtaatacga ctcactatag ggcgaattgg gccctctaga tgcatgctcg agcggccgcc
     5341 agtgtgatgg atatctgcag aattcagg
//
