LOCUS       sequence-386888-        4174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with Arabidopsis ANAC032 CDS and c-terminal 
            HiBiT tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4174)
  AUTHORS   Cai YM, Witham S, Patron NJ
  TITLE     Tuning Plant Promoters Using a Simple Split Luciferase Method to 
            Assess Transcription Factor-DNA Interactions.
  JOURNAL   ACS Synth Biol. 2023 Nov 17;12(11):3482-3486. doi: 
            10.1021/acssynbio.3c00094. Epub 2023 Oct 19.
  PUBMED    37856867
REFERENCE   2  (bases 1 to 4174)
  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"; doi: 
            "10.1021/acssynbio.3c00094"; journalName: "ACS Synth Biol"; date: 
            "2023-11-17- 17"; volume: "12"; issue: "11"; pages: "3482-3486"
COMMENT     Sequence Label: pEPYCeGM0012
FEATURES             Location/Qualifiers
     source          1..4174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(75..102)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(194..280)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    333..432
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1237..1336)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1448..1467)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1526..1545)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2119..2652)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2841..2846,2847..2863,2864..2869))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3526..4114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4015..4034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag ccaggaagag
       61 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
      121 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
      181 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
      241 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
      301 ctcgcgttaa cgctagcatg gatctcgggc cccaaataat gattttattt tgactgatag
      361 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      421 aaaaagcagg cttcatgatg aaatctgggg ctgatttgca atttccacca ggatttagat
      481 ttcatcctac ggatgaggag ctagtcctca tgtatctctg tcgtaaatgc gcgtcgcagc
      541 cgatccctgc tccgattatc accgaactcg atttgtaccg atatgatcct tgggaccttc
      601 ccgacatggc tttgtacggt gaaaaggagt ggtatttttt ctcaccaaga gatcgaaagt
      661 atccaaacgg ttcaagaccc aaccgtgcag ctggtactgg atattggaaa gctaccggag
      721 ctgataaacc aataggtcgt cctaaaccgg ttggtattaa gaaggctcta gtgttttact
      781 cgggaaaacc tccaaatgga gagaaaacca attggattat gcacgaatac cggctcgctg
      841 acgttgaccg gtcggttcgt aagaaaaaca gtctaagatt ggacgattgg gtattgtgtc
      901 gtatatataa caagaaaggt gtcatcgaga agcgacgaag cgatatcgag gacgggttaa
      961 agcctgtgac tgacacgtgt ccaccggaat ctgtggcgag attgatctcc ggctcggagc
     1021 aagcggtgtc accggaattc acgtgtagca acggtcggtt gagtaatgcc cttgattttc
     1081 cgtttaatta cgtagatgcc atcgccgata acgagattgt gtcacggcta ttgggcggga
     1141 atcagatgtg gtcgacgacg cttgatccac ttgtggttag gcagggaact ttcggttcgg
     1201 tgagcggctg gcggctgttc aagaagatta gctgaaaccc agctttcttg tacaaagttg
     1261 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1321 ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa tctctgatgt
     1381 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1441 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1501 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1561 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1621 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1681 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1741 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1801 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1861 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1921 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1981 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2041 ggtgatttct cacttgataa ccttattttt gacgagggga aattaggccg ggaagccgat
     2101 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
     2161 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
     2221 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
     2281 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
     2341 actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
     2401 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
     2461 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
     2521 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
     2581 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     2641 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     2701 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     2761 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     2821 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
     2881 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
     2941 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
     3001 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
     3061 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
     3121 ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata
     3181 gttctagtgg ttggctacta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3241 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3301 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3361 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3421 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttttc
     3481 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3541 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3601 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3661 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4141 gttcctggcc ttttgctggc cttttgctca catg
//
