LOCUS       sequence-444833-        6254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a yellow chromoprotein (visible light and fluorescence) 
            with a his-tag and an inducible GFP (araBAD-promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6254)
  TITLE     Bacterial expression plasmids for education and teaching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6254)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGLO-Yellow-AZA
FEATURES             Location/Qualifiers
     source          1..6254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1312..1334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1342..1344,1345..1347,1348..2061)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1375..1403)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1998..2018
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     RBS             2178..2189
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2917..2928
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2972..2989
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3037..3054)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3207..3293
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3385..3412
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3431..3522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3523..3591,3592..4383)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3741..3760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4425..4880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4512..4531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4722..4743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4991..5579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5480..5499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5733..5750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5765..5905
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5831..5850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5991..6013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6151..6170
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa ttttgtttaa
     1321 ctttaagaag gagatataca tatggctagc aaaggagaag aacttttcac tggagttgtc
     1381 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     1441 ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac tactggaaaa
     1501 ctacctgttc catggccaac acttgtcact actttctctt atggtgttca atgcttttcc
     1561 cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     1621 gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc
     1681 aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga ttttaaagaa
     1741 gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa tgtatacatc
     1801 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa
     1861 gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     1921 gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac
     1981 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
     2041 atggatgagc tctacaaata atgaattcga gctcggtacc cggggatcct ctagaagcgg
     2101 ccgcgtgagg tcagatgctg aacgtcgggt ctcagatgtt gacagctagc tcagtcctag
     2161 gtataatcgg ctgaaacaaa gaggagaaat cttgatgatg acggcattga cggaaggtgc
     2221 aaaactgttt gagaaagaga tcccgtatat caccgaactg gaaggcgacg tcgaaggtat
     2281 gaaatttatc attaaaggcg agggtaccgg tgacgcgagc gttggtaaga ttgacgcgca
     2341 gtttatctgc accacgggtg acgtcccggt gccgtggagc acgttggtga cgacgctgac
     2401 ttacggtgcc caatgtttcg cgaaatatcc gcgtcacatc gcagacttct tcaaatcctg
     2461 tatgccggaa ggttatgtgc aagagcgcac tattaccttt gagggtgacg gtgtctttaa
     2521 gacccgtgcc gaagtgacct tcgaaaacgg tagcgtttac aaccgtgtca agctgaatgg
     2581 ccagggtttc aaaaaggatg gtcacgttct gggtaagaat ctggaattca acttcacccc
     2641 acactgcctg tacatctggg gcgatcaagc gaatcatggt ctgaaaagcg catttaagat
     2701 catgcacgag attaccggct ccaaagaaga tttcatcgtg gctgatcaca cccagatgaa
     2761 taccccgatt ggcggtggcc ctgtgcacgt tccggaatat catcacatta cctaccatgt
     2821 cacgctgagc aaggatgtta ccgaccatcg cgaccacctg aacatcgtag aggttatcaa
     2881 agcagtggac ctggagacgt atcgctaacg gcaagatttc tcctctttgt ttcagccgat
     2941 tatacctagg actgagctag ctgtcaacat ccaccatcac catcaccatt gagcatgcaa
     3001 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     3061 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     3121 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     3181 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3241 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     3301 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     3361 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     3421 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3541 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3601 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3661 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3721 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3781 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3841 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3901 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3961 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4021 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     4081 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4141 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4201 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4261 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4321 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4381 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     4441 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4501 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4561 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4621 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4681 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4741 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4801 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4861 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5101 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5581 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5641 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5701 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5761 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     5821 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     5881 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     5941 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6001 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc
     6061 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     6121 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     6181 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
     6241 tttgacagct tatc
//
