LOCUS       sequence-436610-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a raspberry 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 6189)
  TITLE     Bacterial expression plasmids for education and teaching
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6189)
  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            "Unpublished"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-Pink-SHE
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      131..719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     620..639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     873..890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    905..1045
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(971..990)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1131..1153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1490..2368)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2395..2679
                     /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     2602..2621
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2706..2728
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(2736..2738,2739..2741,2742..3455)
                     /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(2769..2797)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3392..3412
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      complement(3481..3524)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     RBS             3609..3620
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3627..4301
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             4302..4319
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4366..4383)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4536..4622
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4714..4741
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4760..4851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4852..4920,4921..5712)
                     /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(5070..5089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5754..6189,1..20)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5841..5860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6051..6072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
       61 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      241 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
      301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      781 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      841 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      901 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
      961 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     1021 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     1081 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1141 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
     1381 tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     1441 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     1501 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     1561 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     1621 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     1681 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     1741 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     1801 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     1861 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     1921 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     1981 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     2041 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     2101 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     2161 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     2221 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     2281 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     2341 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     2401 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     2461 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     2521 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     2581 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     2641 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa
     2701 ataattttgt ttaactttaa gaaggagata tacatatggc tagcaaagga gaagaacttt
     2761 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     2821 ctgtcagtgg agagggtgaa ggtgatgcta catacggaaa gcttaccctt aaatttattt
     2881 gcactactgg aaaactacct gttccatggc caacacttgt cactactttc tcttatggtg
     2941 ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc aagagtgcca
     3001 tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg aactacaaga
     3061 cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaaaggta
     3121 ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac tataactcac
     3181 acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc
     3241 gccacaacat tgaagatgga tccgttcaac tagcagacca ttatcaacaa aatactccaa
     3301 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgcccttt
     3361 cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg
     3421 ggattacaca tggcatggat gagctctaca aataatgaat tcggcgtatc acgaggcaga
     3481 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctgggtc tcaagagcaa
     3541 cgtctttcgt cttcaataac aactgacagc tagcctgacc ctaggtataa tgttggtacc
     3601 tactagagaa agaggagaaa tactagatgg cttcctccga agacgttatc aaagagttca
     3661 tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg
     3721 aaggtgaagg tcgtccgtac gaaggcaccc agaccgctaa actgaaagtt accaaaggtg
     3781 gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt
     3841 acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca
     3901 aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct
     3961 ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggcaccaac ttcccgtccg
     4021 acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc
     4081 cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc
     4141 actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg
     4201 cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg
     4261 aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc tcaccatcac catcaccatt
     4321 gacatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     4381 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     4441 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     4501 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4561 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4621 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4681 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4741 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     5401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc
     5761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5941 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     6001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6061 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     6121 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6181 ttttaacaa
//
