LOCUS       sequence-437914-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a magenta 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 6231)
  TITLE     Bacterial expression plasmids for education and teaching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6231)
  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            "Unpublished"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-Magenta-AZA
FEATURES             Location/Qualifiers
     source          1..6231
                     /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             2140..2151
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2949..2966
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3014..3031)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3184..3270
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3362..3389
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3408..3499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3500..3568,3569..4360)
                     /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(3718..3737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4402..4857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4489..4508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4699..4720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4968..5556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5457..5476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5710..5727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5742..5882
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5808..5827)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5968..5990
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6128..6147
                     /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 ggtcagatgc tgaacgtcgg gtctcagatg
     2101 ttgacggcta gctcagtcct aggtacagtc ggctgaaaca aagaggagaa atcttgatga
     2161 tggtgagcaa gggtgaggaa ctgattaaag agaatatgcg catgaaagtt gtcatggaag
     2221 gcagcgtgaa tggtcaccag ttcaaatgca ccggcgaggg tgagggtaac ccgtatatgg
     2281 gcacccaaac gatgcgtatc aaagttgttg agggtggccc gttgccgttt gcgttcgaca
     2341 ttttagcgac gagctttatg tatggctctc gtacgtttat caagtacccg aagggtattc
     2401 cggacttttt caaacaatct tttccagagg gtttcacctg ggagcgcgtg actcgctacg
     2461 aagatggcgg cgtcctgacc gcaacgcagg atacctccct ggaagatggc tgcctggtct
     2521 accacgttca ggtccgtggt gtcaatttcc cgagcaatgg tccggttatg cagaagaaaa
     2581 ccctgggttg ggaagcgaac accgagatga tgtatcctgc agatggtggc ctgcgtggtt
     2641 acacccacat ggcattgaaa ctggacggtg gcggccatct gagctgtagc ttcgtgacca
     2701 cgtatcgttc gaagaaaacg gtcggtaaca tcaaaatgcc gggtgtgtac tacgttgacc
     2761 accgtctgga gcgcattaaa gaagccgaca aagagactta cgtggagcaa catgaagtag
     2821 ccgttgcgcg ttattgtgat ctgccgagca agctgggcca taagctgaac agcggtctgc
     2881 gtagccgcgc tcaggccagc aattccgcgg tcgatggtac cgctggtccg ggtagcacgg
     2941 gtagccgtca ccatcaccat caccattgag catgcaagct tggctgtttt ggcggatgag
     3001 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     3061 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     3121 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     3181 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3241 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     3301 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3361 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3421 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3481 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3541 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3601 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3661 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3721 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3781 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3841 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3901 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3961 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4021 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4081 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4141 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4201 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4261 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4321 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4381 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4441 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4741 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4801 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     4861 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4921 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4981 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5041 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5101 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5161 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5221 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5281 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5341 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5401 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5461 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5521 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5581 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5641 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5701 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     5761 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     5821 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     5881 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5941 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6001 catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa cagtcccccg gccacggggc
     6061 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6121 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6181 cggccacgat gcgtccggcg tagaggatct gctcatgttt gacagcttat c
//
