LOCUS       sequence-268978-        7525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of E. coli Phr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7525)
  AUTHORS   Selby CP, Sancar A
  TITLE     A cryptochrome/photolyase class of enzymes with single-stranded 
            DNA-specific photolyase activity.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Nov 21. 103(47):17696-700.
  PUBMED    17062752
REFERENCE   2  (bases 1 to 7525)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Nov 21. 103(47):17696-700."
COMMENT     Sequence Label: pMal-EcPhr
FEATURES             Location/Qualifiers
     source          1..7525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(729..734,735..750,751..757)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    765..781
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     770..792
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             851..1951
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1808..1817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1925..1948
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1982..1993
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3436..3453)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3436..3452)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3445..3467)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3631..3648)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3801..3887
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3979..4006
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4025..4116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4117..4185,4186..4977)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4335..4354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5148..5736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5637..5656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5890..5907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5922..6062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5988..6007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6148..6170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6164..6355)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6851..6928
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6929..7525,1..486)
                     /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(6948..6967)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
       61 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
      121 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
      181 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
      241 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
      301 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
      361 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat tctcatgttt
      541 gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
      601 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
      661 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
      721 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
      781 atttcacaca ggaaacagcc agtccgttta ggtgttttca cgagcacttc accaacaagg
      841 accatagatt atgaaaactg aagaaggtaa actggtaatc tggattaacg gcgataaagg
      901 ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac
      961 cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg
     1021 ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt
     1081 ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc
     1141 cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat
     1201 ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga
     1261 taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt
     1321 cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta
     1381 cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt
     1441 tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc
     1501 ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga
     1561 caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa
     1621 accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc
     1681 gaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga
     1741 caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg
     1801 tattgccgcc accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat
     1861 gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac
     1921 tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tcggtacccg gccggggatc
     1981 catcgagggt aggcctgaat tcatgactac ccatctggtc tggtttcgcc aggatttacg
     2041 tctgcacgat aatctcgcac tggctgccgc ctgccgcaat tcgtctgcac gcgtgctggc
     2101 gttgtatatc gctacaccac gccagtgggc gacgcataac atgtcgccgc gtcaggctga
     2161 acttatcaat gctcaactga atgggctaca aatagcgctt gcggaaaaag gtattcctct
     2221 attgttccgt gaagtggatg actttgtcgc cagtgtcgaa atagttaaac aggtgtgcgc
     2281 ggaaaacagc gttacccacc tgttttataa ctatcagtat gaagtgaatg agcgggcgcg
     2341 ggatgaggaa gttgaaagag cgctgcgtaa cgtggtgtgt gaaggatttg atgacagcgt
     2401 gatcctgccg cctggcgcgg tgatgaccgg taatcacgag atgtacaaag tctttacgcc
     2461 ttttaagaat gcctggctga aacggctgcg ggaagggatg ccggagtgcg tcgctgcacc
     2521 aaaagttcgt agtagcggat cgatagagcc ctcgccatcc attacgctga attatcctcg
     2581 tcagtctttc gatactgcgc attttccggt ggaagaaaaa gcggcgattg cgcaattacg
     2641 ccagttttgc cagaacggtg ccggagaata tgagcaacaa cgagattttc cggcagtgga
     2701 aggcaccagc cgtttgtcgg ccagcctggc aacgggcggg ttgtcgcctc gccagtgctt
     2761 gcatcgcttg ttggctgaac agccgcaggc gctggacggt ggggccggta gtgtctggct
     2821 taatgagctg atctggcgcg agttttaccg tcacctgata acgtatcacc cctcgttgtg
     2881 taaacatcgt ccatttattg cctggacgga tcgtgtacag tggcagagca atcccgcaca
     2941 tttacaggcc tggcaggaag gcaaaacggg atacccgatt gttgatgccg ctatgcgtca
     3001 gcttaacagc actggctgga tgcataacag gctacggatg attacagcca gttttctggt
     3061 gaaagattta ttgatcgact ggcgcgaagg cgagcgatat ttcatgtcgc agctgattga
     3121 tggcgatttg gcagccaata acggtggctg gcagtgggcc gcttcaaccg gaaccgatgc
     3181 agcgccgtat tttcgtattt tcaacccgac aacccaaggc gagaaatttg atcatgaggg
     3241 cgagtttatc cgccagtggc taccggaact gcgcgatgtg ccagggaaag tggtgcatga
     3301 gccgtggaag tgggcgcaga aagcaggtgt gacgctggat tatccgcaac cgatagtcga
     3361 gcacaaagaa gcgagagtac aaacgttggc agcgtatgag gcggcgcgga aggggaaact
     3421 gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3481 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3541 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcagcttgg
     3601 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     3661 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     3721 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     3781 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3841 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     3901 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     3961 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     4021 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4321 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4621 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4981 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5161 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5221 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5281 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5341 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5401 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5461 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5521 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5581 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5641 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5701 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5761 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5821 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5881 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     5941 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     6001 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     6061 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6121 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6181 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6241 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6301 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     6361 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     6421 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     6481 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     6541 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     6601 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     6661 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     6721 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     6781 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     6841 ccgaaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     6901 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     6961 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     7021 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     7081 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     7141 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     7201 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     7261 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     7321 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     7381 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     7441 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     7501 tgtctcggcg cgtctgcgtc tggct
//
