LOCUS       sequence-495030-        5344 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  Final circuit with -Ara: ON, +Ara: OFF behaviour.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 886 to 920)
  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 Dec 10, 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-Y11
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..311
                     /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     234..253
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    372..391
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    498..517
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        886..920
                     /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    1064..1083
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             1088..1110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1118..1831
                     /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      2540..2567
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2755..3570)
                     /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(2755..3570)
                     /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"
     primer_bind     3478..3497
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3571..3662)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3680..4315)
                     /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(3825..3844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4466..5344)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
       61 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
      121 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
      181 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
      241 agcattttta tccataagat tagcggttcc tacctgacgc tttttatcgc aactctctac
      301 tgtttctcca taccgaattc ataggataga ttctggaaac tttaccgtcc gagctccagc
      361 ctgcggtccg ggttcactgc cgtataggca gtctcaagct agactctagt ggtttcagag
      421 ctatgctgga aacagcatag caagttgaaa taaggctagt ccgttatcaa cttgaaaaag
      481 tggcaccgag tcggtgcgtt cactgccgta taggcagtcg ctgggacgcc cgctcgagca
      541 ataaacagtt gatagggctt ctccgttacc atggttcagc caaaaaactt aagaccgccg
      601 gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc ggcggttttc ttttctcttc
      661 tcaattcttc tgacctgtaa cgaataatag atagtaaagt agtctccgat tgagttttct
      721 ctgccgagtc ccacccagtt ctgtgatttc agtaagttgg taattgatac actgttgcga
      781 gaactggtgc ctggtagtag ataggttgtt attgagtaag aaggtaaagt gaacgaaatc
      841 cctgaaactg agactgtaga aaataagctt cagcctgcgg tccggttgac ggctagctca
      901 gtcctaggta cagtgctagc tcgctgggac gcccgagata gccgttacac aggtgacact
      961 tatttcagcc tgcggtccgg ccacactaga gtctagcttg agatcgctgg gacgcccgta
     1021 gaactctgtg aggataaagt ctcccttaca gcctgcggtc cgggttcact gccgtatagg
     1081 cagtaatttt gtttaacttt aagaaggaga tatacatatg cgtaaaggcg aagaactgtt
     1141 taccggtgtg gttccgattc tggtggaact ggacggcgat gttaatggtc ataaattcag
     1201 tgttcgcggc gaaggtgaag gcgatgcgac gaacggcaaa ctgaccctga aatttatctg
     1261 caccacgggt aaactgccgg tcccgtggcc gacgctggtg accacgctga cctatggcgt
     1321 tcaatgtttt gcgcgttacc cggatcacat gaaacagcac gactttttca aatcggccat
     1381 gccggaaggc tatgtgcagg aacgtacgat tagctttaaa gacgatggta cgtataaaac
     1441 ccgcgcggaa gtgaaattcg aaggcgatac cctggttaac cgtatcgaac tgaaaggtat
     1501 cgatttcaaa gaagacggca atattctggg tcataaactg gaatataact tcaattccca
     1561 caacgtgtac atcaccgcgg ataaacagaa aaacggcatt aaagccaatt tcaaaatccg
     1621 ccataatgtg gaagatggta gcgttcagct ggccgaccac tatcagcaaa acacgccgat
     1681 tggtgatggc ccggtcctgc tgccggacaa tcactacctg agtacccagt ccgtgctgtc
     1741 aaaagatccg aacgaaaaac gtgaccacat ggtcctgctg gaatttgtga cggctgcggg
     1801 tatcacccac ggcatggacg aactgtataa aatgtcccgc cgtaatactg acgccatcac
     1861 aatccacagc atcctggatt ggattgaaga ctaataatcg ctgggacgcc cgggatccaa
     1921 gagatttcta cacgattgag cactgtctcc tgcaggctcg gtaccaaatt ccagaaaaga
     1981 ggcctcccga aaggggggcc ttttttcgtt ttggtcctaa tagataaagg ataggtctgg
     2041 tagtgttgtt cgttctcgca ggtaaatcaa taatactctg cagttccgta gacttttcag
     2101 tgggacaggg tagcgataac agatagattg taataagaca cagtaggtgc tcgtagttgc
     2161 gtgaagagaa ccgctcagga aatccagtca gaagtattgg taatcgttga aaactcagtc
     2221 gacgcactta ctgaagacgt cctattacac tcgtcgttgg aaactgaaga tgcggccgcg
     2281 gaaacacaga aaaaagcccg cacctgacag tgcgggcttt ttttttcgac caaaggtagc
     2341 gaacgacgag tcactgttga ggataaatac tttctctact aggcgcctgt tacacaggtc
     2401 ctcagcggcg cgcctttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     2461 gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     2521 gccaggcatc aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttcagatc
     2581 taccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     2641 caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct
     2701 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     2761 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2821 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2881 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2941 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     3001 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     3061 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     3121 agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     3181 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     3241 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     3301 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     3361 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     3421 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     3481 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     3541 gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat
     3601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3661 aataggcatg ctagcgcaga aacgtcctag aagatgccag gaggatactt agcagagaga
     3721 caataaggcc ggagcgaagc cgtttttcca taggctccgc ccccctgacg aacatcacga
     3781 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3841 tccccctgat ggctccctct tgcgctctcc tgttcccgtc ctgcggcgtc cgtgttgtgg
     3901 tggaggcttt acccaaatca ccacgtcccg ttccgtgtag acagttcgct ccaagctggg
     3961 ctgtgtgcaa gaaccccccg ttcagcccga ctgctgcgcc ttatccggta actatcatct
     4021 tgagtccaac ccggaaagac acgacaaaac gccactggca gcagccattg gtaactgaga
     4081 attagtggat ttagatatcg agagtcttga agtggtggcc taacagaggc tacactgaaa
     4141 ggacagtatt tggtatctgc gctccactaa agccagttac caggttaagc agttccccaa
     4201 ctgacttaac cttcgatcaa accgcctccc caggcggttt tttcgtttac agagcaggag
     4261 attacgacga tcgtaaaagg atctcaagaa gatcctttac ggattcccga caccatcact
     4321 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     4381 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4441 ctcctagggt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     4501 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     4561 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     4621 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     4681 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     4741 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     4801 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     4861 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     4921 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     4981 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     5041 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     5101 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     5161 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     5221 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     5281 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     5341 ccat
//
