LOCUS       sequence-451049-        4412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mEOS3.2-PSP-GSS-FLAG-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4412)
  AUTHORS   Castanzo DT, LaFrance B, Martin A
  TITLE     The AAA+ ATPase Msp1 is a processive protein translocase with robust
            unfoldase activity.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jun 30;117(26):14970-14977. doi: 
            10.1073/pnas.1920109117. Epub 2020 Jun 15.
  PUBMED    32541053
REFERENCE   2  (bases 1 to 4412)
  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"; doi: 
            "10.1073/pnas.1920109117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-06-30- 30"; volume: "117"; issue: "26"; pages: 
            "14970-14977"
COMMENT     Sequence Label: pAM253
FEATURES             Location/Qualifiers
     source          1..4412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(196..213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(330..875)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(410..429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1100..2182)
                     /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     2144..2163
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2183..2260)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2306..2325
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2306..2324
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2325..2349
                     /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)."
     CDS             2393..3070
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             3071..3094
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3104..3127
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3128..3145
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3172..3190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3186..3233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3454..4113)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4028..4047
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4114..4216)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat
       61 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
      121 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
      181 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
      241 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
      301 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
      361 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
      421 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
      481 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
      541 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
      601 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
      661 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
      721 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
      781 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
      841 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
      901 aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
      961 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     1021 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     1081 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     1201 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     1261 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     1321 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     1381 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     1441 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     1501 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     1561 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     1621 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     1681 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     1741 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     1801 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     1861 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     1921 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     1981 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     2041 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     2101 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     2161 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     2221 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     2281 ttatgcgact cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat
     2341 aacaattccc ctgtagaaat aattttgttt aactttaata aggagatata ccatgagtgc
     2401 gattaagcca gacatgaaga tcaaactccg tatggaaggc aacgtaaacg ggcaccactt
     2461 tgtgatcgac ggagatggta caggcaagcc ttttgaggga aaacagagta tggatcttga
     2521 agtcaaagag ggcggacctc tgccttttgc ctttgatatc ctgaccactg cattccatta
     2581 cggcaacagg gtattcgcca aatatccaga caacatacaa gactatttta agcagtcgtt
     2641 tcctaagggg tattcgtggg aacgaagctt gactttcgaa gacgggggca tttgcaacgc
     2701 cagaaacgac ataacaatgg aaggggacac tttctataat aaagttcgat tttatggtac
     2761 caactttccc gccaatggtc cagttatgca gaagaagacg ctgaaatggg agccctccac
     2821 tgagaaaatg tatgtgcgtg atggagtgct gacgggtgat attgagatgg ctttgttgct
     2881 tgaaggaaat gcccattacc gatgtgactt cagaactact tacaaagcta aggagaaggg
     2941 tgtcaagtta ccaggcgccc actttgtgga ccactgcatt gagattttaa gccatgacaa
     3001 agattacaac aaggttaagc tgtatgagca tgctgttgct cattctggat tgcctgacaa
     3061 tgccagacga ctggaagtcc tgttccaagg tccgggcagc agcgattata aggatgacga
     3121 tgacaagcat caccatcatc accactgatt aattaaccta ggctgctgcc accgctgagc
     3181 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac
     3241 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3301 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     3361 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaaccagg
     3421 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
     3481 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     3541 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatagt
     3601 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     3661 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     3721 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     3781 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     3841 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaactc
     3901 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     3961 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     4021 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     4081 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     4141 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     4201 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     4261 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc
     4321 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
     4381 gaggtgctcc agtggcttct gtttctatca gc
//
