LOCUS       sequence-218821-        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"
     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(2390..3268)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3295..3579
                     /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     3502..3521
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             3589..3597
                     /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             3603..4319
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTRVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKVRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     4256..4276
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4515..4534)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        4601..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4693..5502
                     /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 tatgacaact
     2401 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     2461 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     2521 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     2581 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     2641 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     2701 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     2761 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     2821 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     2881 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     2941 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     3001 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     3061 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     3121 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     3181 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     3241 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     3301 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     3361 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     3421 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     3481 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     3541 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgttttta aggaggtaaa
     3601 aaatgagtaa aggagaagaa cttttcacta gagttgtccc aattcttgtt gaattagacg
     3661 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg agaaggtgat gcaacatacg
     3721 gaaaacttac ccttaaattt atttgcacta ctggaaaact accagttccg tggccaacac
     3781 ttgtcactac tctctcttat ggtgttcaat gcttttcgag atacccagat cacatgaaac
     3841 ggcatgactt tttcaagagt gccatgcccg agggttatgt acaggaaaga actatatttt
     3901 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
     3961 ttaatagaat cgagttaaaa ggtgttgatt ttaaagaaga tggaaacatt cttggacaca
     4021 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     4081 gaatcaaagc taacttcaaa gttagacaca acattgaaga tggaagcgtt caactggcag
     4141 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     4201 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     4261 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataac
     4321 cccaagggcg acacccccta attagcccgg gcgaaaggcc cagtctttcg actgagcctt
     4381 tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagtcccc acactaccat
     4441 cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac
     4501 tgccgccagg caaacaaggg gtgttatgag ccatattcag gtataaatgg gctcgcgata
     4561 atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga atatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     4741 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     4801 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     4861 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     4921 atgccacttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     4981 actgcgatcc ccggaaaaac agcgttccag gtattagaag aatatcctga ttcaggtgaa
     5041 aatattgttg atgcgctggc agtgttcctg cgccggttgc actcgattcc tgtttgtaat
     5101 tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg aatgaataac
     5161 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     5221 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     5281 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     5341 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     5401 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     5461 aataaattgc aatttcattt gatgctcgat gagtttttct aagcggcgcg ccatcgaatg
     5521 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     5581 gaat
//
