LOCUS       sequence-324478-        8072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBP-Anacystis nidulans photolyase in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8072)
  TITLE     A. nidulans photolyase-MBP fusion  protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8072)
  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            "Unpublished"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: pMalAnPL
FEATURES             Location/Qualifiers
     source          1..8072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             123..1223
                     /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      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1197..1220
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1272..1283
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2760..2777)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2760..2776)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2769..2791)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2955..2972)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3125..3211
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3303..3330
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3349..3440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3441..3509,3510..4301)
                     /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(3659..3678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4343..4856)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4538..4559)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4750..4769
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4967..5555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5456..5475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5709..5726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5741..5881
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5807..5826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5967..5989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5983..6174)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6670..6747
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6748..7830
                     /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(6767..6786)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaac tgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
      181 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
      241 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
      301 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
      361 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
      421 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
      481 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
      541 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
      601 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
      661 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
      721 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
      781 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
      841 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
      901 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt
      961 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     1021 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     1081 ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     1141 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     1201 aagccctgaa agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca
     1261 acaacctcgg gatcgaggga aggatttcag aattcatggc ggctccgatt ctgttttggc
     1321 accggcgtga cctgcgcctc agcgacaaca ttggtctggc ggcggcgcgg gcacaatcag
     1381 cgcagctgat cggtctgttt tgcctcgacc ctcagattct ccagagtgct gacatggcgc
     1441 cagcacgggt tgcctacctg cagggctgcc tacaggaact ccagcagcgt tatcaacagg
     1501 cgggcagtcg gctgctattg ctccaagggg atccacaaca cctgatcccg caactggcgc
     1561 agcagcttca ggcggaagca gtttattgga accaagacat tgaaccctac ggccgcgatc
     1621 gcgatgggca ggtcgcagcc gctctgaaaa cggcaggcat tcgggctgtt caactctggg
     1681 atcaactgct ccatagcccc gatcaaatcc tcagcggcag cggtaatccc tacagcgtct
     1741 acgggccgtt ttggaaaaat tggcaggctc agcccaagcc cacgccggtt gccacaccga
     1801 cagaactggt cgatctctca ccagagcaac taactgcgat cgcacctctg ctgttgtcgg
     1861 aactaccgac cctgaagcag cttggctttg actgggacgg gggttttcca gtggagccgg
     1921 gggaaacagc ggcgatcgcc cgtcttcaag aattttgcga tcgcgcgatc gccgactacg
     1981 atccccagcg caattttccg gcggaagcag gaacctcggg cttgagtccc gctttgaaat
     2041 tcggggcgat cggtattcgt caggcttggc aggctgccag tgcagcccat gccctcagcc
     2101 gcagcgacga agcccgcaac agcattcgcg tttggcagca ggaactcgct tggcgggagt
     2161 tctatcagca tgcgctctac catttcccga gcctggccga tggtccctat cgctcgctct
     2221 ggcagcaatt cccttgggaa aatcgcgagg ctctgttcac ggcttggacc caggcgcaaa
     2281 cgggctatcc gattgttgat gcagcgatgc gccaactgac tgaaaccggc tggatgcaca
     2341 accgttgccg gatgattgtt gccagctttt tgaccaagga tttgatcatc gactggcggc
     2401 gtggggagca gttctttatg cagcacttgg tcgatggcga tctggctgca aataacggcg
     2461 gttggcagtg gagcgcttcc agtggcatgg atcccaaacc gctgcggatt ttcaatcctg
     2521 ctagtcaggc caagaagttt gatgccacgg cgacctacat caaacgttgg ctgccagaac
     2581 tgcgccatgt ccatcccaag gacttgatca gtggggagat cacgccgatt gaacgccgag
     2641 gctatcctgc cccgatcgtg aatcacaacc tgcgccagaa acagttcaag gcgctctaca
     2701 accagctcaa agcggctatc gctgaacctg aggctgagcc cgattcatag aagcttggca
     2761 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2821 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2881 ccttcccaac agttgcgcag cctgaatggc gaatggcagc ttggctgttt tggcggatga
     2941 gataagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     3001 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     3061 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     3121 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3181 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     3241 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     3301 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3421 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3601 tccttgagag ttttcgcccc gaagaacgtt tcccaatgat gagcactttt aaagttctgc
     3661 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3961 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4321 catatatact ttagattgat ttaccccggt tgataatcag aaaagcccca aaaacaggaa
     4381 gattgtataa gcaaatattt aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     4441 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4501 atcaaaagaa tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4561 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4621 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4681 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4741 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4801 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaaa
     4861 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4921 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4981 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5041 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5101 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5161 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5221 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5281 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5341 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5401 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5461 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5521 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5581 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5641 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5701 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5761 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     5821 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     5881 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5941 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6001 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     6061 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     6121 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     6181 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     6241 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     6301 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     6361 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     6421 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     6481 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     6541 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     6601 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     6661 cgaaattccg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     6721 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     6781 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     6841 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     6901 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     6961 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     7021 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     7081 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     7141 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     7201 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     7261 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     7321 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     7381 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     7441 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     7501 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     7561 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     7621 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     7681 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     7741 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7801 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     7861 tcattaggca caattctcat gtttgacagc ttatcatcga ctgcacggtg caccaatgct
     7921 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
     7981 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat
     8041 aacggttctg gcaaatattc tgaaatgagc tg
//
