LOCUS       sequence-173033-        7633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CBD and intein tag containing AbaSI enzyme full length.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  AUTHORS   Kazrani AA, Kowalska M, Czapinska H, Bochtler M
  TITLE     Crystal structure of the 5hmC specific endonuclease PvuRts1I.
  JOURNAL   Nucleic Acids Res. 2014 May;42(9):5929-36. doi: 10.1093/nar/gku186. 
            Epub 2014 Mar 14.
  PUBMED    24634440
REFERENCE   2  (bases 1 to 7633)
  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: 
            "10.1093/nar/gku186"; journalName: "Nucleic Acids Res"; date: 
            "2014-05"; volume: "42"; issue: "9"; pages: "5929-36"
COMMENT     Sequence Label: pTXB1-AbaSFI
FEATURES             Location/Qualifiers
     source          1..7633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      86..172
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      269..355
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      452..538
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      635..721
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     856..875
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    875..899
                     /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             914..936
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1922..2515
                     /codon_start=1
                     /gene="Mycobacterium xenopi gyrA"
                     /product="modified GyrA intein (Southworth et al., 1999)"
                     /label=Mxe GyrA intein
                     /note="N-terminal cleavage of this intein can be induced by
                     thiols."
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             2546..2701
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(2766..2784)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2780..2827
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2887..2991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2992..3060,3061..3852)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3210..3229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3894..4407)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4089..4110)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4301..4320
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4518..5106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5007..5026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5260..5277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5299..5318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5459..5481
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5475..5666)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6223..7305)
                     /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     7267..7286
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7306..7383)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      7536..7622
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
       61 tgccaggaat taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
      121 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
      181 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
      241 aactgccagg aattaattcc ccaggcatca aataaaacga aaggctcagt cgaaagactg
      301 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
      361 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
      421 ataaactgcc aggaattaat tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      481 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
      541 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
      601 gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg ctcagtcgaa
      661 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
      721 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
      781 cccgccataa actgccagga attggggatc ggaattaatt cccggtttaa accggggatc
      841 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      901 ctctagaaat aattttgttt aactttaaga aggagatata catatgtgca ataaggcgtc
      961 gagcgatctt acggattacg ttattcgcca gttgggccgt acaaaaaata aacgctatga
     1021 agcctatgtt gtaagccgca tcattcatct tctgaatgac ttcaccctca aattcgtcac
     1081 ccagcaattc gtacgcctgt ctaataagaa aattgcattg actgatctgt atttcccgca
     1141 actgggtatc cacatcgaag tggatgaagg ccatcacttt ctgcgcaata gcaaaatgga
     1201 atacagtctg aaccaaattg atgagccgct ttactccatt tcacagacag agagtgatgc
     1261 tatgcgtgaa gaagatatca ttagcattac tggccacaaa atctttcgcg ttaatgtgtt
     1321 caagaatcag gaagggcaac cacagaactt agaaaacatc catcagcaga ttgacaaaat
     1381 cattgaggag atcaaaactg ccaagaacaa actcattgaa gcgtcaacct ttaaagagtg
     1441 gaacattgaa accgaatata atccgcagac gtatatcgat ctgggccgta tttccttagc
     1501 agacaatgtg gtcctcaaaa ctacgaaaga tgtgtgtaac tgctttgggt atagttacaa
     1561 gaactaccaa cgcggtggtg ccttacatcc ctacaagaaa gacacactga tttggtttcc
     1621 acggctgtat gagaacaaag actggattaa cacgatttcc ccggatggac tgacgatcac
     1681 cgagaaaagc acggatgaaa ccatcacctt gaagaagctg gaagaatgga agaatggccc
     1741 tcagaaacgc attgtgtttg cgcgtgtgaa agataacctg tcgtcgcgtg cgatgtaccg
     1801 gtttatggga ctgtatgaat tccagaaagc agacctgaaa gatggtgctg tttggaaacg
     1861 ggtcaaatgc gaagtacaaa cgtattctcc gaaagaaacc aaatgtctcg agggctcttc
     1921 ctgcatcacg ggagatgcac tagttgccct acccgagggc gagtcggtac gcatcgccga
     1981 catcgtgccg ggtgcgcggc ccaacagtga caacgccatc gacctgaaag tccttgaccg
     2041 gcatggcaat cccgtgctcg ccgaccggct gttccactcc ggcgagcatc cggtgtacac
     2101 ggtgcgtacg gtcgaaggtc tgcgtgtgac gggcaccgcg aaccacccgt tgttgtgttt
     2161 ggtcgacgtc gccggggtgc cgaccctgct gtggaagctg atcgacgaaa tcaagccggg
     2221 cgattacgcg gtgattcaac gcagcgcatt cagcgtcgac tgtgcaggtt ttgcccgcgg
     2281 gaaacccgaa tttgcgccca caacctacac agtcggcgtc cctggactgg tgcgtttctt
     2341 ggaagcacac caccgagacc cggacgccca agctatcgcc gacgagctga ccgacgggcg
     2401 gttctactac gcgaaagtcg ccagtgtcac cgacgccggc gtgcagccgg tgtatagcct
     2461 tcgtgtcgac acggcagacc acgcgtttat cacgaacggg ttcgtcagcc acgctactgg
     2521 cctcaccggt ctgaactcag gcctcacgac aaatcctggt gtatccgctt ggcaggtcaa
     2581 cacagcttat actgcgggac aattggtcac atataacggc aagacgtata aatgtttgca
     2641 gccccacacc tccttggcag gatgggaacc atccaacgtt cctgccttgt ggcagcttca
     2701 atgactgcag gaaggggatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2761 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2821 ttttttgctg aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga
     2881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3181 ttcccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     3241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg
     3901 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     3961 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4021 aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga
     4081 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4141 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     4201 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4261 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4321 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4381 cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat
     4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5161 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5221 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5281 gctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     5341 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     5401 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5461 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     5521 taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc atccgcgtcc
     5581 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     5641 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     5701 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     5761 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     5821 cagagaaaaa tcactcaggg tcaatgccag ccgaacgcca gcaagacgta gcccagcgcg
     5881 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     5941 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     6001 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     6061 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     6121 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     6181 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     6241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6301 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     6361 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     6421 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     6481 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     6541 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     6601 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     6661 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     6721 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     6781 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     6841 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6901 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6961 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     7021 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     7081 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     7141 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     7201 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     7261 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     7321 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     7381 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     7441 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgccggcat gccgcccttt
     7501 cgtcttcaag aattaattcc caattcccca ggcatcaaat aaaacgaaag gctcagtcga
     7561 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     7621 atccgccggg agc
//
