LOCUS       sequence-387962-        5014 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Level 0 Tp901 integrase with GG1 and GG4 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  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 5014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-I42 Tp901 GG1-GG4
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1615..1631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1615..1631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1628..1650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1639..1655
                     /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(1663..1669,1670..1687,1688..1693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1708..1729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1846..1863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2017..2605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2743..3117)
                     /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      3114..3123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3323..4117)
                     /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(3323..4117)
                     /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(3434..3453)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4044..4063
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(4466..4768)
                     /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(4568..4587)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     4891..4913
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4905..4922
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4906..4922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4929..4948
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4929..4947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggatcggagg tctcaaatga ctaagaaagt agcaatctat acacgagtat ccactactaa
       61 ccaagcagag gaaggcttct caattgatga gcaaattgac cgtttaacaa aatatgctga
      121 agcaatgggg tggcaagtat ctgatactta tactgatgct ggtttttcag gggccaaact
      181 tgaacgccca gcaatgcaaa gattaatcaa cgatatcgag aataaagctt ttgatacagt
      241 tcttgtatat aagctagacc gcctttcacg tagtgtaaga gatactcttt atcttgttaa
      301 ggatgtgttc acaaaaaata aaatagactt tatctcgctt aatgaaagta ttgatacttc
      361 ttctgctatg ggtagcttgt ttctcactat tctttctgca attaatgagt ttgaaagaga
      421 gaatataaaa gaacgcatga ctatgggtaa actagggcga gcgaaatctg gtaagtctat
      481 gatgtggact aagacagctt ttgggtatta ccacaacaga aagacaggta tattagaaat
      541 tgttccttta caagctacaa tagttgaaca aatattcact gattatttat caggaatatc
      601 acttacaaaa ttaagagata aactcaatga atctggacac atcggtaaag atataccgtg
      661 gtcttatcgt accctaagac aaacacttga taatccagtt tactgtggtt atatcaaatt
      721 taaggacagc ctatttgaag gtatgcacaa accaattatc ccttatgaga cttatttaaa
      781 agttcaaaaa gagctagaag aaagacaaca gcagacttat gaaagaaata acaaccctag
      841 acctttccaa gctaaatata tgctgtcagg gatggcaagg tgcggttact gtggagcacc
      901 tttaaaaatt gttcttggcc acaaaagaaa agatggaagc cgcactatga aatatcactg
      961 tgcaaataga tttcctcgaa aaacaaaagg aattacagta tataatgaca ataaaaagtg
     1021 tgattcagga acttatgatt taagtaattt agaaaatact gttattgaca acctgattgg
     1081 atttcaagaa aataatgact ccttattgaa aattatcaat ggcaacaacc aacctattct
     1141 tgatacttcg tcatttaaaa agcaaatttc acagatcgat aaaaaaatac aaaagaactc
     1201 tgatttgtac ctaaatgatt ttatcactat ggatgagttg aaagatcgta ctgattccct
     1261 tcaggctgag aaaaagctgc ttaaagctaa gattagcgaa aataaattta atgactctac
     1321 tgatgttttt gagttagtta aaactcagtt gggctcaatt ccgattaatg aactatcata
     1381 tgataataaa aagaaaatcg tcaacaacct tgtatcaaag gttgatgtta ctgctgataa
     1441 tgtagatatc atatttaaat tccaactcgc taccggttaa taagctttga gacctcatca
     1501 cgcctgaatt ccagcacact ggcggccgtt actagtggat ccgagctcgg taccaagctt
     1561 gatgcatagc ttgagtattc taacgcgtca cctaaatagc ttggcgtaat catggtcata
     1621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2701 cacctagatc cttttaaatt aaaaatgaag ttttagcacg tgtcagtcct gctcctcggc
     2761 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
     2821 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
     2881 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
     2941 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
     3001 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
     3061 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
     3121 ggccctcctc acgtgctatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3241 agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3301 ggaaattgta agcgttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc
     3361 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     3421 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     3481 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     3541 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     3601 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     3661 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     3721 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     3781 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     3841 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     3901 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     3961 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     4021 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     4081 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     4141 cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     4201 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     4261 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     4321 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     4381 ggggtcagca ccgtttctgc ggactggctt tctacgtgaa aaggatctag gtgaagatcc
     4441 tttttgataa tctcatgcct gacatttata ttccccagaa catcaggtta atggcgtttt
     4501 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
     4561 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
     4621 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
     4681 gtcgccccgg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     4741 ctcttttata ggtgtaaacc ttaaactgcc gtacgtatag gctgcgcaac tgttgggaag
     4801 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4861 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4921 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     4981 gccgccagtg tgatggatat ctgcagaatt cagg
//
