LOCUS       sequence-493106-        5344 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Final circuit with -Ara: ON, +Ara: ON behaviour.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 2978 to 3012)
  AUTHORS   Nevot G, Guell M, Santos-Moreno J
  TITLE     Critical Analysis of Preprints and Inquiry-Based Lessons Improve the
            Synthetic Biology Learning Experience.
  JOURNAL   ACS Synth Biol. 2025 Aug 15;14(8):2878-2884. doi: 
            10.1021/acssynbio.5c00014.
  PUBMED    40814254
REFERENCE   2  (bases 1 to 5344)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/acssynbio";
            journalName: "ACS Synth Biol"; date: "2025-08-15- 15"; volume: "14";
            issue: "8"; pages: "2878-2884"
COMMENT     Sequence Label: pEND-Z11
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     226..245
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(319..410)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(428..1063)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(573..592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1214..2092)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2119..2403
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2326..2345
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    2464..2483
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    2590..2609
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        2978..3012
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     protein_bind    3156..3175
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             3180..3202
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3210..3923
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4632..4659
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4847..5344,1..318))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(join(4847..5344,1..318))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
       61 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
      121 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
      181 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
      241 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
      301 ccgttgaata tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgct
      421 agcgcagaaa cgtcctagaa gatgccagga ggatacttag cagagagaca ataaggccgg
      481 agcgaagccg tttttccata ggctccgccc ccctgacgaa catcacgaaa tctgacgctc
      541 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctgatgg
      601 ctccctcttg cgctctcctg ttcccgtcct gcggcgtccg tgttgtggtg gaggctttac
      661 ccaaatcacc acgtcccgtt ccgtgtagac agttcgctcc aagctgggct gtgtgcaaga
      721 accccccgtt cagcccgact gctgcgcctt atccggtaac tatcatcttg agtccaaccc
      781 ggaaagacac gacaaaacgc cactggcagc agccattggt aactgagaat tagtggattt
      841 agatatcgag agtcttgaag tggtggccta acagaggcta cactgaaagg acagtatttg
      901 gtatctgcgc tccactaaag ccagttacca ggttaagcag ttccccaact gacttaacct
      961 tcgatcaaac cgcctcccca ggcggttttt tcgtttacag agcaggagat tacgacgatc
     1021 gtaaaaggat ctcaagaaga tcctttacgg attcccgaca ccatcactct agatttcagt
     1081 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     1141 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgct cctagggtct
     1201 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     1261 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     1321 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     1381 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     1441 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     1501 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     1561 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     1621 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     1681 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     1741 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     1801 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     1861 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     1921 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     1981 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     2041 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     2101 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
     2161 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
     2221 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
     2281 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
     2341 tatccataag attagcggtt cctacctgac gctttttatc gcaactctct actgtttctc
     2401 cataccgaat tcataggata gattctggaa actttaccgt ccgagctcca gcctgcggtc
     2461 cgggttcact gccgtatagg cagatcagtg tgtactaagt actgtttcag agctatgctg
     2521 gaaacagcat agcaagttga aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     2581 agtcggtgcg ttcactgccg tataggcagt cgctgggacg cccgctcgag caataaacag
     2641 ttgatagggc ttctccgtta ccatggttca gccaaaaaac ttaagaccgc cggtcttgtc
     2701 cactaccttg cagtaatgcg gtggacagga tcggcggttt tcttttctct tctcaattct
     2761 tctgacctgt aacgaataat agatagtaaa gtagtctccg attgagtttt ctctgccgag
     2821 tcccacccag ttctgtgatt tcagtaagtt ggtaattgat acactgttgc gagaactggt
     2881 gcctggtagt agataggttg ttattgagta agaaggtaaa gtgaacgaaa tccctgaaac
     2941 tgagactgta gaaaataagc ttcagcctgc ggtccggttg acggctagct cagtcctagg
     3001 tacagtgcta gctcgctggg acgcccgaga tagccgttac acaggtgaca cttatttcag
     3061 cctgcggtcc ggccacacta gagtctagct tgagatcgct gggacgcccg tagaactctg
     3121 tgaggataaa gtctccctta cagcctgcgg tccgggttca ctgccgtata ggcagtaatt
     3181 ttgtttaact ttaagaagga gatatacata tgcgtaaagg cgaagaactg tttaccggtg
     3241 tggttccgat tctggtggaa ctggacggcg atgttaatgg tcataaattc agtgttcgcg
     3301 gcgaaggtga aggcgatgcg acgaacggca aactgaccct gaaatttatc tgcaccacgg
     3361 gtaaactgcc ggtcccgtgg ccgacgctgg tgaccacgct gacctatggc gttcaatgtt
     3421 ttgcgcgtta cccggatcac atgaaacagc acgacttttt caaatcggcc atgccggaag
     3481 gctatgtgca ggaacgtacg attagcttta aagacgatgg tacgtataaa acccgcgcgg
     3541 aagtgaaatt cgaaggcgat accctggtta accgtatcga actgaaaggt atcgatttca
     3601 aagaagacgg caatattctg ggtcataaac tggaatataa cttcaattcc cacaacgtgt
     3661 acatcaccgc ggataaacag aaaaacggca ttaaagccaa tttcaaaatc cgccataatg
     3721 tggaagatgg tagcgttcag ctggccgacc actatcagca aaacacgccg attggtgatg
     3781 gcccggtcct gctgccggac aatcactacc tgagtaccca gtccgtgctg tcaaaagatc
     3841 cgaacgaaaa acgtgaccac atggtcctgc tggaatttgt gacggctgcg ggtatcaccc
     3901 acggcatgga cgaactgtat aaaatgtccc gccgtaatac tgacgccatc acaatccaca
     3961 gcatcctgga ttggattgaa gactaataat cgctgggacg cccgggatcc aagagatttc
     4021 tacacgattg agcactgtct cctgcaggct cggtaccaaa ttccagaaaa gaggcctccc
     4081 gaaagggggg ccttttttcg ttttggtcct aatagataaa ggataggtct ggtagtgttg
     4141 ttcgttctcg caggtaaatc aataatactc tgcagttccg tagacttttc agtgggacag
     4201 ggtagcgata acagatagat tgtaataaga cacagtaggt gctcgtagtt gcgtgaagag
     4261 aaccgctcag gaaatccagt cagaagtatt ggtaatcgtt gaaaactcag tcgacgcact
     4321 tactgaagac gtcctattac actcgtcgtt ggaaactgaa gatgcggccg cggaaacaca
     4381 gaaaaaagcc cgcacctgac agtgcgggct ttttttttcg accaaaggta gcgaacgacg
     4441 agtcactgtt gaggataaat actttctcta ctaggcgcct gttacacagg tcctcagcgg
     4501 cgcgcctttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     4561 cgttgtgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     4621 tcaaactaag cagaaggcca tcctgacgga tggccttttt gcgtttcaga tctaccggta
     4681 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gacaagctga
     4741 cgaccgggtc tccgcaagtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4801 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     4861 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     4921 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     4981 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     5041 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     5101 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     5161 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     5221 gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     5281 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     5341 ggaa
//
