LOCUS       sequence-7396-ex        6154 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6154)
  AUTHORS   Lee JY, Kwak JE, Moon J, Eom SH, Liong EC, Pedelacq JD, Berendzen J,
            Suh SW
  TITLE     Crystal structure and functional analysis of the SurE protein 
            identify a novel phosphatase family.
  JOURNAL   Nat Struct Biol. 2001 Sep . 8(9):789-94.
  PUBMED    11524683
REFERENCE   2  (bases 1 to 6154)
  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: "Nat Struct 
            Biol. 2001 Sep . 8(9):789-94."
COMMENT     Sequence Label: pET-21a(+) SurE
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             958..980
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    995..1019
                     /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     complement(1019..1038)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1020..1038)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1122..1141)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1347..1424
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1425..2507
                     /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(1444..1463)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3316..3507
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3501..3523)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3609..3751
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3664..3683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3766..3783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3937..4525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4017..4036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4696..5487,5488..5556))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5319..5338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5557..5661)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5688..6143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5825..5846)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6037..6056
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tcctcagtcg tacacctccc ttaacttctt cagacactgt
      241 tcgttcgtga gaaagggatg tatcggagtg atggagacgt acccttctct aaccgccttg
      301 tagtcaacat cgtctctatc gtcgtcttcg atgacctctc ccatcatcca gtagtacttc
      361 tccccaaagg gtgagactct ctcttcgaag taatcgttcc accttcttct gctctgcctt
      421 gtgaatctcc atccctttat ctcaccagca ggcacgttta tgttcagcat ggtgaaagga
      481 tcgaggagtg agaagtcgaa ttctttcaga aagtcgatca aaaaacgtgc cgctccttcg
      541 aaatcgggac tctcgtagtt agcactcgat atggcaatcg aaggaatgtt catcatggca
      601 ccttccatcg cacccgagac cgtcccagag tggagaatat ccatgcccat gttgggcccc
      661 ctgttcacac cactcacgat gagatcgacc cttttatcca tgaccacgtt gtaggcgagt
      721 ttcacacagt ccgcgggcgt tcctgtagtg gagtacgcaa ccaccctctc tgagatgaaa
      781 accttcttca tccagagcgg aacgtgtatg gtgatgctgt gaccagttgc gctcctctct
      841 ttatcgggag ccaccacgaa cacctcgtgt tcctcggaaa gaagctctgc gaggacgatg
      901 atccccttag actgtatccc gtcggcgttg gttaccagaa ttctcatcat atgtatatct
      961 ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc
     1021 ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat
     1081 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     1141 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     1201 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     1261 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     1321 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
     1381 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     1441 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     1501 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     1561 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     1621 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     1681 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     1741 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     1801 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     1861 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     1921 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     1981 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     2041 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     2101 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     2161 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     2221 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     2281 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     2341 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     2401 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2461 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2521 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
     2581 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2641 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
     2701 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     2761 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     2821 caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg
     2881 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     2941 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     3001 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
     3061 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     3121 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
     3181 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
     3241 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
     3301 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     3361 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
     3421 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
     3481 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3541 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3601 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     3661 atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg aaataccgca
     3721 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt
     5701 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5761 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5821 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5881 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5941 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6001 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6061 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6121 taatgcgccg ctacagggcg cgtcccattc gcca
//
