LOCUS       sequence-212515-        5533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP from jellyfish Aquorea sp. under D61 AND gate-like ara+lac 
            promoter with AraC repressor; lacI repressor; kanamycin resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5533)
  AUTHORS   Siddall A, Williams AA, Sanders J, Denton JA, Madden D, Schollar J, 
            Bryk J
  TITLE     UNIGEMS: plasmids and parts to facilitate teaching on assembly, gene
            expression control and logic in E. coli
  JOURNAL   bioRxiv 2021.06.20.449138
REFERENCE   2  (bases 1 to 5533)
  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: "bioRxiv 
            2021.06.20.449138"
COMMENT     Sequence Label: p006-GFP-Logic-AND
FEATURES             Location/Qualifiers
     source          1..5533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      476..1064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     965..984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1218..1235
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1334..1350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1703..2581)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     protein_bind    2800..2816
                     /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)."
     CDS             2859..3575
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTRVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKVRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     3512..3532
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3771..3790)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3857..3948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3949..4758
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(4841..5533,1..390)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4860..4879)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
       61 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
      121 ggatacgacg ataccgaaga tagctcatgt tatatcccgc cgttaaccac catcaaacag
      181 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
      241 gcggtgaagg gcaatcagct gttgccagtc tcactggtga aaagaaaaac caccctggcg
      301 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      361 caggtttccc gactggaaag cgggcagtga ctcatgacca aaatccctta acgtgagtta
      421 cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      601 gagcgcagat accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga
      661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1141 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1201 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac
     1261 tgccaggcat caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa
     1321 ctctttctgt gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga
     1381 taacgagtaa tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg
     1441 gggggagttt agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg
     1501 atatatgaga attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata
     1561 cgttgctttt tcgattgatg aacacctata attaaactat tcatctatta tttatgattt
     1621 tttgtatata caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata
     1681 ataaagggaa aatcagttat acttatgaca acttgacggc tacatcattc actttttctt
     1741 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     1801 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     1861 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     1921 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     1981 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     2041 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     2101 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     2161 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     2221 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     2281 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     2341 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     2401 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     2461 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     2521 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     2581 tattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     2641 tgcgccattc gatgggcttc ctaatggaat acgatgacat gtggatgggc agtggccggt
     2701 tgttacacgc ctaccgcgat gctgaatgac ccggactaaa gtggcgaaaa ttatgtacaa
     2761 cgtcgtgtta gctgcctttt agcaatttta tccatagact tgtgagcgga taacaatgat
     2821 acttgaatac atctggcggt gataaggcgt tacccaacat gagtaaagga gaagaacttt
     2881 tcactagagt tgtcccaatt cttgttgaat tagacggtga tgttaatggg cacaaatttt
     2941 ctgtcagtgg agagggagaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     3001 gcactactgg aaaactacca gttccgtggc caacacttgt cactactctc tcttatggtg
     3061 ttcaatgctt ttcgagatac ccagatcaca tgaaacggca tgactttttc aagagtgcca
     3121 tgcccgaggg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     3181 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggtg
     3241 ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac
     3301 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaagtta
     3361 gacacaacat tgaagatgga agcgttcaac tggcagacca ttatcaacaa aatactccaa
     3421 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     3481 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
     3541 ggattacaca tggcatggat gaactataca aataacccca agggcgacac cccctaatta
     3601 gcccgggcga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt
     3661 tccctactct cgcatgggga gtccccacac taccatcggc gctacggcgt ttcacttctg
     3721 agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa caaggggtgt
     3781 tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt
     3841 gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3901 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagccatatt
     3961 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     4021 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     4081 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     4141 gatgagatgg tcagactaaa ctggctgacg gaatttatgc cacttccgac catcaagcat
     4201 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagcg
     4261 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     4321 ttcctgcgcc ggttgcactc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     4381 tttcgcctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     4441 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     4501 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     4561 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     4621 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     4681 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     4741 ctcgatgagt ttttctaagc ggcgcgccat cgaatggcgc aaaacctttc gcggtatggc
     4801 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgaaaccag taacgttata
     4861 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4921 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4981 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     5041 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     5101 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     5161 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     5221 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     5281 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aggacggtac
     5341 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     5401 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     5461 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     5521 acaaaccatg caa
//
