LOCUS       sequence-218819-        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"
     CDS             1..1083
                     /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(20..39)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1169..1757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1658..1677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1911..1928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2027..2043
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2396..3274)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     protein_bind    3493..3509
                     /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             3552..4268
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTRVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKVRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     4205..4225
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4464..4483)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        4550..4641
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4642..5451
                     /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"
ORIGIN
        1 atgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
       61 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
      121 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
      181 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
      241 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
      301 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
      361 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
      421 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
      481 ttctcccatg aggacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
      541 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
      601 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
      661 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      721 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      781 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agatagctca
      841 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      901 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgcca
      961 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     1021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1081 tgactcatga ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag
     1141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1321 tagtgtagcc gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg
     1381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1861 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1921 tgagcgagga agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc
     1981 ccctgacgga tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc
     2041 agtcttaagc tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata
     2101 acgtaaaaac ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt
     2161 cagaatattt aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa
     2221 atgagaaaaa agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct
     2281 ataattaaac tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt
     2341 taaagagaat taagaaaata aatctcgaaa ataataaagg gaaaatcagt tatacttatg
     2401 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
     2461 tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat
     2521 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
     2581 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
     2641 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt
     2701 ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat
     2761 ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg
     2821 ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt
     2881 cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg
     2941 acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat
     3001 accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca
     3061 aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc
     3121 gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga
     3181 gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc
     3241 ccggcagcag gggatcattt tgcgcttcag ccatattcac caccctgaat tgactctctt
     3301 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatgggc ttcctaatgg
     3361 aatacgatga catgtggatg ggcagtggcc ggttgttaca cgcctaccgc gatgctgaat
     3421 gacccggact aaagtggcga aaattatgta caacgtcgtg ttagctgcct tttagcaatt
     3481 ttatccatag acttgtgagc ggataacaat gatacttgaa tacatctggc ggtgataagg
     3541 cgttacccaa catgagtaaa ggagaagaac ttttcactag agttgtccca attcttgttg
     3601 aattagacgg tgatgttaat gggcacaaat tttctgtcag tggagaggga gaaggtgatg
     3661 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta ccagttccgt
     3721 ggccaacact tgtcactact ctctcttatg gtgttcaatg cttttcgaga tacccagatc
     3781 acatgaaacg gcatgacttt ttcaagagtg ccatgcccga gggttatgta caggaaagaa
     3841 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
     3901 atacccttgt taatagaatc gagttaaaag gtgttgattt taaagaagat ggaaacattc
     3961 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
     4021 aaaagaatgg aatcaaagct aacttcaaag ttagacacaa cattgaagat ggaagcgttc
     4081 aactggcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     4141 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
     4201 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     4261 acaaataacc ccaagggcga caccccctaa ttagcccggg cgaaaggccc agtctttcga
     4321 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcatgg ggagtcccca
     4381 cactaccatc ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca
     4441 ccgcgctact gccgccaggc aaacaagggg tgttatgagc catattcagg tataaatggg
     4501 ctcgcgataa tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt
     4561 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4621 taatattgaa aaaggaagaa tatgagccat attcaacggg aaacgtcgag gccgcgatta
     4681 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     4741 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     4801 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     4861 acggaattta tgccacttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     4921 ttactcacca ctgcgatccc cggaaaaaca gcgttccagg tattagaaga atatcctgat
     4981 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ctcgattcct
     5041 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga
     5101 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     5161 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     5221 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     5281 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     5341 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     5401 cctgatatga ataaattgca atttcatttg atgctcgatg agtttttcta agcggcgcgc
     5461 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     5521 cagggtggtg aat
//
