LOCUS       sequence-345506-        6757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His14-bdSUMO-AwaIscB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6757)
  AUTHORS   Altae-Tran H, Kannan S, Demircioglu FE, Oshiro R, Nety SP, McKay LJ,
            Dlakic M, Inskeep WP, Makarova KS, Macrae RK, Koonin EV, Zhang F
  TITLE     The widespread IS200/IS605 transposon family encodes diverse 
            programmable RNA-guided endonucleases.
  JOURNAL   Science. 2021 Oct;374(6563):57-65. doi: 10.1126/science.abj6856. 
            Epub 2021 Sep 9.
  PUBMED    34591643
REFERENCE   2  (bases 1 to 6757)
  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: 
            "10.1126/science.abj6856"; journalName: "Science"; date: "2021-10"; 
            volume: "374"; issue: "6563"; pages: "57-65"
COMMENT     Sequence Label: pHS0806 His14-bdSUMO-AwaIscB
FEATURES             Location/Qualifiers
     source          1..6757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     992..1011
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        992..1010
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1011..1035
                     /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)."
     RBS             1050..1072
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2796..2814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2810..2857
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2894..3349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2981..3000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3191..3212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3464..4255,4256..4324))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4087..4106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4325..4417)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4498..5086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4987..5006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5240..5257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5272..5411
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5337..5356)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5497..5519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5513..5704)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6276..6757,1..601))
                     /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"
ORIGIN
        1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     1021 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     1081 gagcaagcat caccatcatt caggccatca ccataccgga caccaccatc attcaggcag
     1141 tcatcaccat accggatctg ctgcgggtgg cgaagaagat aagaaaccgg caggtggcga
     1201 aggtggcggt gcccatatca acctgaaagt gaaaggtcaa gacggcaacg aagtcttttt
     1261 ccgcatcaaa cgttctaccc agctgaaaaa gctgatgaac gcatactgtg accgtcagtc
     1321 tgtagacatg accgcaattg ctttcctgtt tgatggtcgt cgcctgcgtg cggaacagac
     1381 cccggatgaa ctggagatgg aagatggcga cgaaatcgac gcaatgcttc atcagactgg
     1441 tggctcaagt ggatccatga gcgtgttcgt gctgaacaag cagaaacggc ctctgatgcc
     1501 ctgcagcgag aagagagccc gcctgctgct ggaaagaggc agagccgtgg tggtgcggct
     1561 ggtgccattc accatccggc tgaaggatag aatcggcggc gtgctgcagc ccctgagact
     1621 gaagctggac cctggctcta agacaaccgg catcgccctg gtgcgcgagg tggtgcggcg
     1681 ggacgagagc gtggtttggc tggctgaact gacacacaga ggatatcaga tcagcgaggc
     1741 cctgagagct agaagcgcca tgagaagaag aagaaggtcc gccaacctgc ggtacagagc
     1801 ccctagattc ctgaatcgga ccaagcctaa gggctggctg gcccctagcc tgagacatag
     1861 agtggaaacc accattaact gggtcaagag actgcggaga ctggctccta tcacggaaat
     1921 cacccaggag ctggtcaggt tcgacctgca agccatgcag caccccgaga tcagcggcat
     1981 tgagtaccag cagggcgagc tggctggcta cgaggtgcgg gaatacctgc tggaaaagtg
     2041 gcagcggaca tgcgcctact gcggcgccca gcaggtgccc ctccagatcg agcacatcag
     2101 acctaagtcc gccggcggaa gcaacagatt gtctaatctc accctggctt gcgccccttg
     2161 caaccacaag aagggcgctc aatctatcga ggccttcctg aagcacaaac tggaactgct
     2221 gaaacagatc caggcccagg cacaagcccc tctgaaagac gccgccgccg tgaacaccac
     2281 ccggtgggcc ctgttcaacg ccctgaaggc tacaggcctg caggtgaaaa ccggcagcgg
     2341 cggacagacc aaatacaacc gccagagact gggcatccca aagacccacg ccctggacgc
     2401 cgcttgtgtg ggcaagctgg atgccctgca caactggcag atccctacac tggccatcaa
     2461 ggcgatgggc agaggcagct accagaggac cagactgaat agattcggct tccccagagg
     2521 ccacctgatg agacacaaga gaatccacgg ttttcagacc ggcgacagag tgatcgcaca
     2581 catccccagc ggcaaaaagg ccggagtgca cgtgggaaga gtggccgtcc gaaccagcgg
     2641 atcttttaac atccaaacag ccacaggcgt gatccagggc atcgcccacc ggcattgtag
     2701 cgtcctgcag cgggccgacg gctacggcta cagcttcaac ctgacccagc ctgaggaagc
     2761 cagactggct gcctgattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
     2821 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2881 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3241 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3301 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc tggcggcacg
     3361 atggcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3421 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3481 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3541 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3601 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3661 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3721 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3781 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3841 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3901 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3961 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4021 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4081 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4141 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4201 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4261 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4321 tcatactctt cctttttcaa tcatgattga agcatttatc agggttattg tctcatgagc
     4381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gtcatgacca aaatccctta
     4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4621 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     5341 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     5401 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5461 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5521 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5581 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5641 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5701 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     5761 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     5821 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     5881 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     5941 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     6001 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     6061 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     6121 gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg ccccgcgccc
     6181 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     6241 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6301 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6361 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     6421 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     6481 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     6541 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     6601 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     6661 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     6721 atcggctgaa tttgattgcg agtgagatat ttatgcc
//
