LOCUS       sequence-212478-        5584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP from jellyfish Aquorea sp. under pBAD promoter with AraC 
            repressor; kanamycin resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  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 5584)
  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-pBAD
FEATURES             Location/Qualifiers
     source          1..5584
                     /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(1697..2575)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2602..2886
                     /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     2809..2828
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2896..2904
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2910..3626
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTRVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKVRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     3563..3583
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3822..3841)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3908..3999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4000..4809
                     /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(4892..5584,1..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(4911..4930)
                     /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 gacaacttga cggctacatc attcactttt tcttcacaac
     1741 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     1801 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     1861 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     1921 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     1981 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     2041 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     2101 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     2161 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     2221 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     2281 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     2341 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     2401 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     2461 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     2521 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     2581 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     2641 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     2701 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     2761 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     2821 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     2881 ctccataccc gtttttaagg aggtaaaaaa tgagtaaagg agaagaactt ttcactagag
     2941 ttgtcccaat tcttgttgaa ttagacggtg atgttaatgg gcacaaattt tctgtcagtg
     3001 gagagggaga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     3061 gaaaactacc agttccgtgg ccaacacttg tcactactct ctcttatggt gttcaatgct
     3121 tttcgagata cccagatcac atgaaacggc atgacttttt caagagtgcc atgcccgagg
     3181 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     3241 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt gttgatttta
     3301 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     3361 acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaagtt agacacaaca
     3421 ttgaagatgg aagcgttcaa ctggcagacc attatcaaca aaatactcca attggcgatg
     3481 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     3541 ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     3601 atggcatgga tgaactatac aaataacccc aagggcgaca ccccctaatt agcccgggcg
     3661 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3721 tcgcatgggg agtccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     3781 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa acaaggggtg ttatgagcca
     3841 tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg
     3901 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3961 ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagccatat tcaacgggaa
     4021 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     4081 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     4141 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     4201 gtcagactaa actggctgac ggaatttatg ccacttccga ccatcaagca ttttatccgt
     4261 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc gttccaggta
     4321 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     4381 cggttgcact cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc
     4441 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     4501 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     4561 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     4621 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     4681 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     4741 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     4801 tttttctaag cggcgcgcca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     4861 cccggaagag agtcaattca gggtggtgaa tatgaaacca gtaacgttat acgatgtcgc
     4921 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     4981 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5041 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5101 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5161 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5221 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5281 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5341 ctctgaccag acacccatca acagtattat tttctcccat gaggacggta cgcgactggg
     5401 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     5461 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     5521 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     5581 gcaa
//
