LOCUS       sequence-7406-ex        6312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  AUTHORS   Kwak JE, Lee JY, Han BW, Moon JH, Sohn SH, Park IS, Kim BG, Suh SW
  TITLE     Crystallization and preliminary X-ray crystallographic analysis of 
            RNase HIII from Bacillus subtilis.
  JOURNAL   Acta Crystallogr D Biol Crystallogr. 2001 Mar . 57(Pt 3):438-40.
  PUBMED    11223525
REFERENCE   2  (bases 1 to 6312)
  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: "Acta 
            Crystallogr D Biol Crystallogr. 2001 Mar . 57(Pt 3):438-40."
COMMENT     Sequence Label: pET-22b(+)-RNase HIII
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1116..1138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1153..1177
                     /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(1177..1196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1178..1196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1280..1299)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1505..1582
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1583..2665
                     /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(1602..1621)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3474..3665
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3659..3681)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3767..3909
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3822..3841
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3924..3941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4095..4683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4175..4194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4854..5645,5646..5714))
                     /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     5477..5496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5715..5819)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5846..6301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5983..6004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6195..6214
                     /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 gagctatgaa cgttttttat
      181 cagcaaggcg ctgcgctttt tgcgtattgg cgaaatggag ttttgtaaag gttctgagtg
      241 cagatgcccc ttttttgagg atcagcttgg cggccgcttc atcgacgtgc ggaccggccc
      301 cttttggaag cgtcattcct gccgcgcgtg atagtttatc catttccatt aaaaatgaat
      361 atcttgcaat gattgaagca gcggcgacag ccaggtgaat accttcagct tttgtgctaa
      421 aataggtccg ctccttcaca atgtctctgc cctttaaatg attgaagtag acgcctggct
      481 cagcgaattg gtcaatcaaa atggcttcag gctttactcc atccaatttt cttaataaat
      541 gtgtgatcgc ctgattgtgc agcaaagctt tcattttccc ttggctcatg cctttttcct
      601 gcatgctgtt atacttttca ttctttaaca ccagcaggct gtacggaatg gttttaatca
      661 ggtttcgggc aatctcgatg atttgcggat cttttaaatc tttagaatcc ttaacaccga
      721 gttccttcat taaaggaagc atggttttgt cgacgtacgc gcatacaacc gtcattgggc
      781 caaagtaatc tccggttccg acttcgtcag aaccgataac agacattccg gcgatatctg
      841 ccggaggggc atagcgggga tcagccggct ttttggccgt tttcttcttt tcctgaggct
      901 ctgctgttcc ccagcgcgcg gattctgccg cagcgttttt tccttgaaac aagaccttac
      961 ctgattgata tgctgtaatg gtacagccgg gcggttttgc ctgaaaaacg gctccctgcg
     1021 gaacagaggc tgtaagtgaa ccgctgtacg tcattttcat ttggtcaata gcagacaacg
     1081 atacttttat cactgaatgg gacaccatat gtatatctcc ttcttaaagt taaacaaaat
     1141 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     1201 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     1261 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     1321 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     1381 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     1441 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     1501 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     1561 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1621 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1681 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1741 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     1801 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     1861 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     1921 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     1981 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     2041 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     2101 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     2161 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     2221 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     2281 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     2341 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     2401 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     2461 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     2521 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     2581 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2641 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     2701 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2761 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2821 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2881 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     2941 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     3001 cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     3061 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     3121 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     3181 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     3241 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     3301 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     3361 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     3421 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     3481 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     3541 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     3601 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     3661 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3721 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     3781 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     3841 actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3901 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4501 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4981 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5161 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5821 acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt aaaattcgcg
     5881 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5941 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     6001 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     6061 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6121 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6181 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6241 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6301 tcccattcgc ca
//
