LOCUS       sequence-177284-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Srap1 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  AUTHORS   Kweon SM, Zhu B, Chen Y, Aravind L, Xu SY, Feldman DE
  TITLE     Erasure of Tet-Oxidized 5-Methylcytosine by a SRAP Nuclease.
  JOURNAL   Cell Rep. 2017 Oct 10;21(2):482-494. doi: 
            10.1016/j.celrep.2017.09.055.
  PUBMED    29020633
REFERENCE   2  (bases 1 to 6005)
  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: 
            "10.1016/j.celrep.2017.09"; journalName: "Cell Rep"; date: 
            "2017-10-10- 10"; volume: "21"; issue: "2"; pages: "482-494"
COMMENT     Sequence Label: pGEX4T-1 Srap1
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(530..535,536..553,554..560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    568..584
                     /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     592..608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     592..608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             604..1131
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(624..641)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(624..640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(633..655)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1233..1238,1239..1254,1255..1261)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1269..1285
                     /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             1308..1961
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1919..1941
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1968..1985
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3105..3127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3272..3290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3358..3462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3463..3531,3532..4323)
                     /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(3681..3700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4494..5082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4983..5002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5236..5253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5326..5403
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5404..6005,1..481)
                     /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(5423..5442)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
       61 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
      121 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
      181 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
      241 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
      301 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
      361 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct ttacacttta
      541 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      601 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      661 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      721 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      781 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      841 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      901 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
      961 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     1021 acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct
     1081 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
     1141 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat
     1201 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
     1261 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac
     1321 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     1381 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     1441 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     1501 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     1561 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     1621 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     1681 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     1741 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     1801 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     1861 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     1921 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gttccgcgtg
     1981 gatcctgcgg gcgaacgtcc tgtcacttgc ccagagaggt tctcaccagg gcctgcgcct
     2041 atcaggatcg gcagggccgg cggcggctcc cgcagtggag ggaccccgac aagtactgcc
     2101 cctcctacaa caagagcccg cagtccagca gccccgtgct gctctccaga ctgcactttg
     2161 agaaggatgc agactcatca gatcggataa ttattcccat gcgatggggc ttagtcccat
     2221 cttggttcaa agaaagtgat ccttctaagc tgcagttcaa cactaccaac tgtcgcagtg
     2281 ataccataat ggagaagcag tcattcaagg ttcctctggg gaaaggacgg cggtgtgttg
     2341 ttttagcaga tggattctac gagtggcagc ggtgtcaggg aacaaaccag aggcaaccat
     2401 acttcatcta ttttcctcaa atcaagacag agaagtcagg tgggaacgat gcttcagaca
     2461 gctctgacaa caaggaaaag gtctgggaca actggaggct gctgacaatg gcagggatct
     2521 ttgactgctg ggaagcgcca gggggagagt gcctgtattc ctacagcatc atcactgtgg
     2581 attcctgcag aggtttgagt gacatccaca gcaggatgcc tgccatacta gatggagaag
     2641 aagcagtctc caaatggctc gactttggtg aggtcgccac tcaggaagct ctgaagctaa
     2701 tccaccccat agacaatatc accttccatc cagtttctcc agtggtgaac aattcccgaa
     2761 acaacactcc ggagtgtctg gcgcctgctg acttgctggt taagaaggag cccaaggcaa
     2821 atggcagcag tcaaaggatg atgcagtggc tggctacaaa gtcacccaaa aaggaagtcc
     2881 ctgactcacc caaaaaggat gcatcaggtc taccccagtg gtccagccag tttctccaga
     2941 agagcccatt gcctgctaaa agaggtgcta ccagcagctt cctggatcga tggctgaagc
     3001 aggagaagga ggatgagccc atggccaaga agcctaacag ctgagcggcc gcatcgtgac
     3061 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     3121 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3181 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     3241 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3301 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3361 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3421 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3481 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3541 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3601 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3661 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3721 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3781 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3841 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3901 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3961 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     4021 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4081 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4141 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4201 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4261 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4321 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4381 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4621 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5161 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5221 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     5281 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     5341 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     5401 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     5461 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     5521 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     5581 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     5641 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     5701 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     5761 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     5821 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     5881 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     5941 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     6001 ggctg
//
