LOCUS       sequence-386872-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pARF19 promoter with GG0 and GG1 golden gate linkers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  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 5550)
  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: pNL2114 pARF19 GG0-GG1
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2105..2124)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2106..2124)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2131..2148)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2131..2147)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2140..2162)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2285..2587
                     /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     2466..2485
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2936..3730
                     /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             2936..3730
                     /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(2990..3009)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3600..3619
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3930..3939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3936..4310
                     /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      4448..5036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4937..4956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5190..5207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5324..5345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5360..5365,5366..5383,5384..5390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5398..5414
                     /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     5403..5425
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5422..5438
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5422..5438
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg tctcaggagc cacgcgcagc aaatattgac ccaaaaaggc ataaaaaaca
       61 ctaaagcatg ccaacttttt attcttgttg agatttactt tgtcgtccca tatcttttca
      121 aattttgatc ctctataata tatgtttaaa cctaacttat caaactttac attccttttt
      181 cttttgctca ctaggtaagg ctcttgagat cgcttcacct catctcttgt ttaacattta
      241 taaacatgta gctgtctggt taatcaattt acatgtggta aattataaat tactctagaa
      301 agagatgtat attaaactgc taagttcatg gctaatcttc ggtatttctt gtttaaccgc
      361 gataagttga acatatatta atgagctctt atggtttttg tttaatgtcc atttggaggt
      421 gtgatcatct catatgttct caccaaaaga ttaccacttg aattctttta catgtgaaat
      481 gttttaattt attaaccctc ttctgttttg atcaaaacat aaactatata tttgaatctt
      541 ttatatataa ttcatattaa ttaaataaca tcatgttttc aatgaatgta ttgacatgat
      601 atcaatgaag aatatttact ttggaaagag tcagagttgt tctcgaaaga tatttaaatg
      661 ttccaaatta tttccgtttg tctaaacata gaaaatccct aattacatat aaatcaacaa
      721 aaaggtaata attaggtcaa aacctccccc actaaataca tgtatactga gattcagaaa
      781 tacaatacta ttatagtata tgttaattga aagcaaaatt cataagaaag atcggcgttg
      841 aaaggaaaaa ggctctacaa ctacatttta tatgcatacg tagagagctg aaagtaagga
      901 acaactctac tccggtccag attaattgta aagaacttta tacattaatt aaaacaaaaa
      961 cgaacacttt atactattga ttaaaaaaag aagaagatga ataaaataat tgcctgaaaa
     1021 attaataatt acacatgaag gcttagcttt aattggagta taatgatgtc cgtagctctg
     1081 taaacatgtg aagttctcct ccttttgcgg ctagtgggtg ctccattgcg tatcaaaagg
     1141 aatattattc taatcataat taagtcatta aattactagt agtgtttttt tttttctcgc
     1201 attatacgga catgtctcaa tttggaccaa tccatataaa tgaataaact tattagctac
     1261 tactatatta taacagaaaa tatgtatata aaagaaaaaa agtgggacat cttttctggt
     1321 aaccaagagt ttacaacttt ggattagaaa atatttaagt ccccatttgg cttgcaattt
     1381 tcggtccgcc gaacctaatc cttaggtttt tctcaatata tccgggttta tggtataacc
     1441 tcattcataa gaaaatccgt attactaaag taaaaaatta aaaatatggt tataaaaacg
     1501 agggaaagtt aacagatttt aaaaccatgg actgtttttc caacaacagt aaattataat
     1561 cggaaaagat aaaaataaaa agaaataaaa aaggtgggac cgacaagaag aagacatcaa
     1621 gtggggtgcc acgtcagaag gcataagaaa agatttcatc tttccgcatg tatccgatcc
     1681 caatcggctt aaatctcaat atcgattatt gtttttattt ttttgaaaaa acacttgaga
     1741 atctgcagaa acacgagaaa aaaaagaaaa gtttcaaagc tttttaagag cttaaaaatt
     1801 tgctttgaag cttcaaatat tcttatgaac taaaaagaag aaaaaagctt ttgtttcttt
     1861 ttccttagca gcagaatgat ttttgtttcc aaaattatta ctatttagtt tctctcgtgc
     1921 tcttctcttg agcaaataca gattcgttaa ttttgctgaa gaagaagaac tctgtttctt
     1981 ccctgcacca aaccaatttt ttcgttcttt ctataaacca atgtgagacc acgaagtgcc
     2041 tgaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccaa
     2101 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     2161 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2221 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt
     2281 acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     2341 acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg
     2401 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     2461 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     2521 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     2581 aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta
     2641 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     2701 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     2761 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     2821 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     2881 ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat
     2941 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3001 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3061 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3121 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3181 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3241 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3301 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3361 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3421 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3481 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3541 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3601 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3661 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3721 gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     3781 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3841 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3901 aaatgcttca ataatagcac gtgaggaggg ccaccatggc caagttgacc agtgccgttc
     3961 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     4021 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4081 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4141 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4201 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4261 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctaa
     4321 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4381 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4441 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4501 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4561 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4621 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4681 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4741 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4801 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4861 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4921 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4981 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5041 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5101 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5161 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5221 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5281 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5341 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5401 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctattta
     5461 ggtgacgcgt tagaatactc aagctatgca tcaagcttgg taccgagctc ggatccacta
     5521 gtaacggccg ccagtgtgct ggaattcagg
//
