LOCUS       Exported                7684 bp ds-DNA     circular SYN 12-JUL-2024
DEFINITION  Cloning vector pTXB3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7684)
  AUTHORS   Telenti A, Southworth M, Alcaide F, Daugelat S, Jacobs WR Jr., 
            Perler FB.
  TITLE     The Mycobacterium xenopi GyrA protein splicing element: 
            characterization of a minimal intein
  JOURNAL   J Bacteriol 179, 6378-6382 (1997)
  PUBMED    9335286
REFERENCE   2  (bases 1 to 7684)
  AUTHORS   Evans TC Jr., Benner J, Xu M-Q.
  TITLE     Semisynthesis of cytotoxic proteins using a modified protein 
            splicing element
  JOURNAL   Protein Sci 7, 2256-2264 (1998)
  PUBMED    9827992
REFERENCE   3  (bases 1 to 7684)
  AUTHORS   Southworth MW, Amaya K, Evans TC, Xu M-Q., Perler FB.
  TITLE     Purification of proteins fused to either the amino or carboxy 
            terminus of the Mycobacterium xenopi Gyrase A intein
  JOURNAL   BioTechniques 27, 110-120 (1999)
  PUBMED    10407673
REFERENCE   4  (bases 1 to 7684)
  AUTHORS   Perler FB.
  TITLE     InBase, the intein database
  JOURNAL   Nucleic Acids Res 30 (1), 383-384 (2002)
  PUBMED    11752343
REFERENCE   5  (bases 1 to 7684)
  AUTHORS   Ghosh I, Xu MQ.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2007) Research Department, New England Biolabs, 
            240 County Road, Ipswich, MA 01938, USA
REFERENCE   6  (bases 1 to 7684)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 15, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          join(1..5724,6771..7684)
                     /organism="Cloning vector pTXB3"
                     /mol_type="other DNA"
                     /label=source:Cloning vector pTXB3
                     /note="/locus_tag=source:Cloning vector pTXB3"
     gene            140..1000
                     /gene="bla"
                     /locus_tag="bla"
                     /label=bla
     CDS             140..1000
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="beta-lactamase"
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     sig_peptide     140..208
                     /gene="bla"
                     /locus_tag="bla(1)"
                     /label=bla
                     /label=bla(1)
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein"
     rep_origin      1042..1555
                     /locus_tag="M13 origin of replication (+ -)"
                     /label=M13 origin of replication (+ -)
                     /note="M13 origin of replication (+ -)"
     rep_origin      1666..2254
                     /locus_tag="pMB1 origin of replication (clockwise) (RNAII
                     -35 to RNA/DNA switch point)"
                     /label=pMB1 origin of replication (clockwise)
                     (RNAII -...
                     /label=pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point)
                     /note="pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point)"
     gene            complement(2623..2814)
                     /gene="rop"
                     /locus_tag="rop"
                     /label=rop
     CDS             complement(2623..2814)
                     /codon_start=1
                     /transl_table=11
                     /exception="alternative start codon"
                     /gene="rop"
                     /locus_tag="ROP protein"
                     /product="ROP protein"
                     /label=ROP protein
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(3371..4453)
                     /gene="lacI"
                     /locus_tag="lacI"
                     /label=lacI
     CDS             complement(3371..4453)
                     /codon_start=1
                     /transl_table=11
                     /exception="alternative start codon"
                     /gene="lacI"
                     /locus_tag="lac repressor (LacI)"
                     /product="lac repressor (LacI)"
                     /label=lac repressor (LacI)
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5637..5656
                     /locus_tag="T7 universal primer sequence (NEB #S1248S)"
                     /label=T7 universal primer sequence (NEB
                     #S1248S)
                     /label=T7 universal primer sequence (NEB #S1248S)
                     /note="T7 universal primer sequence (NEB #S1248S)"
     promoter        5637..5654
                     /locus_tag="T7 promoter (transcript start 5654 clockwise)"
                     /label=T7 promoter (transcript start 5654
                     clockwise)
                     /label=T7 promoter (transcript start 5654 clockwise)
                     /note="T7 promoter (transcript start 5654 clockwise)"
     protein_bind    5656..5680
                     /locus_tag="lac operator"
                     /label=lac operator
                     /bound_moiety="LacI"
                     /note="lac operator"
     RBS             5710..5716
                     /locus_tag="RBS for expression CDS"
                     /label=RBS for expression CDS
                     /note="RBS for expression CDS"
     misc_feature    5722..5724
                     /gene="expression CDS"
                     /locus_tag="expression CDS"
                     /label=expression CDS
                     /note="multiple cloning site (NdeI-SapI)"
     misc_feature    5725..5727
                     /locus_tag="start codon"
                     /label=start codon
     gene            6771..7536
                     /gene="expression CDS"
                     /locus_tag="expression CDS(1)"
                     /label=expression CDS
                     /label=expression CDS(1)
     CDS             6771..7536
                     /codon_start=1
                     /gene="expression CDS"
                     /locus_tag="intein/CBD fusion"
                     /product="intein/CBD fusion"
                     /label=intein/CBD fusion
                     /note="multiple cloning site (NdeI-SapI) 5722..5775; Mxe
                     gyrA intein sequence 5776..6369; chitin binding domain 
                     (CBD) sequence 6400..6558"
                     /translation="MSCPTRGRVGTHRRHRAGCAAQQ*QRHRPESP*PAWQSRARRPAV
                     PLRRASGVHGAYGRRSACDGHREPPVVVFGRRRRGADPAVEADRRNQAGRLRGDSTQRI
                     QRRLCRFCPRETRICAHNLHSRRPWTGAFLGSTPPRPGRPSYRRRADRRAVLLRESRQC
                     HRRRRAAGV*PSCRHGRPRVYHERVRQPRYWPHRSELRPHDKSWCIRLAGQHSLYCGTI
                     GHI*RQDV*MFAAPHLLGRMGTIQRSCLVAASM"
     primer_bind     complement(6804..6821)
                     /locus_tag="Mxe intein reverse primer sequence (NEB
                     #S1268S, discontinued; replaced by #S1285S)"
                     /label=Mxe intein reverse primer sequence (NEB
                     #S1268S...
                     /label=Mxe intein reverse primer sequence (NEB #S1268S, 
                     discontinued; replaced by #S1285S)
                     /note="Mxe intein reverse primer sequence (NEB #S1268S, 
                     discontinued; replaced by #S1285S)"
     primer_bind     complement(6864..6883)
                     /locus_tag="Mxe intein reverse II primer sequence (NEB
                     #S1285S)"
                     /label=Mxe intein reverse II primer sequence (NEB
                     #S12...
                     /label=Mxe intein reverse II primer sequence (NEB
                     #S1285S)
                     /note="Mxe intein reverse II primer sequence (NEB
                     #S1285S)"
     terminator      7556..7678
                     /locus_tag="T7 Tphi transcription terminator"
                     /label=T7 Tphi transcription terminator
                     /note="T7 Tphi transcription terminator"
ORIGIN
        1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
       61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      301 ccttgagagt tttcgccccg aagaacgttc tccaatgatg agcactttta aagttctgct
      361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
     1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
     1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg
     2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcc
     3001 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     3061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     3661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4501 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4561 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4621 aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca attccccagg
     4681 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     4741 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     4801 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc
     4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc
     5041 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
     5221 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5281 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     5341 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     5401 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     5581 aattggggat cggaattaat tcccggttta aaccggggat ctcgatcccg cgaaattaat
     5641 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5701 taactttaag aaggagatat aaccatggcg tcgaaggcca acgtcggaat caacggtttc
     5761 ggaagaatcg gacgtctcgt gctccgcgcg gctgtcgaaa aggacacggt tcaagtggtc
     5821 gccgtcaacg atccgtttat cacaatcgac tacatggtct accttttcaa gtatgactcg
     5881 acccacggtc aattcaaagg aactgttacc tacgatggag actttctgat tgtccagaaa
     5941 gatggcaaat catcgcacaa gatcaaggtc ttcaacagca aggatccagc tgccatcgca
     6001 tggggatcag tcaaagccga tttcgttgtc gagtctactg gagttttcac gacaaaggaa
     6061 aaggcttctg ctcatcttca aggaggagcc aagaaggtca tcatctctgc tccatctgct
     6121 gatgctccaa tgtacgtggt tggagtaaac catgagaagt acgacgcttc gaatgatcac
     6181 gttgtttcta acgcatcgtg caccaccaac tgtctggcac cactggcgaa ggttatcaat
     6241 gataacttcg gtatcatcga aggactcatg acgacagtgc acgctgtaac cgcaacccag
     6301 aagacagtcg atggaccatc cggaaagctg tggagagatg gacgcggtgc tggacaaaac
     6361 atcattccag cttccactgg agccgccaag gctgtgggaa aagtcattcc ggagctgaac
     6421 ggaaaactca ctggaatggc tttccgtgtc cctacaccag atgtttccgt cgttgatctg
     6481 accgttcgcc tcgagaagcc agcttcgatg gacgacatca agaaggtagt caaggccgct
     6541 gccgatggac caatgaaggg aatcctcgcc tacactgaag atcaagttgt gtcgactgat
     6601 ttcgtgtccg accctcactc gtccattttc gatgctggag catgcatttc gctcaacccg
     6661 aacttcgtca agctcgtctc ttggtacgac aatgaatatg gatactcgaa ccgtgttgtc
     6721 gacctcatcg gatacatcgc cacccgtgga gcatgcatca cgggagatgc actagttgcc
     6781 ctacccgagg gcgagtcggt acgcatcgcc gacatcgtgc cgggtgcgcg gcccaacagt
     6841 gacaacgcca tcgacctgaa agtccttgac cggcatggca atcccgtgct cgccgaccgg
     6901 ctgttccact ccggcgagca tccggtgtac acggtgcgta cggtcgaagg tctgcgtgtg
     6961 acgggcaccg cgaaccaccc gttgttgtgt ttggtcgacg tcgccggggt gccgaccctg
     7021 ctgtggaagc tgatcgacga aatcaagccg ggcgattacg cggtgattca acgcagcgca
     7081 ttcagcgtcg actgtgcagg ttttgcccgc gggaaacccg aatttgcgcc cacaacctac
     7141 acagtcggcg tccctggact ggtgcgtttc ttggaagcac accaccgaga cccggacgcc
     7201 caagctatcg ccgacgagct gaccgacggg cggttctact acgcgaaagt cgccagtgtc
     7261 accgacgccg gcgtgcagcc ggtgtatagc cttcgtgtcg acacggcaga ccacgcgttt
     7321 atcacgaacg ggttcgtcag ccacgctact ggcctcaccg gtctgaactc aggcctcacg
     7381 acaaatcctg gtgtatccgc ttggcaggtc aacacagctt atactgcggg acaattggtc
     7441 acatataacg gcaagacgta taaatgtttg cagccccaca cctccttggc aggatgggaa
     7501 ccatccaacg ttcctgcctt gtggcagctt caatgactgc aggaagggga tccggctgct
     7561 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     7621 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     7681 ggat
//
