LOCUS       sequence-484213-        6923 bp    DNA     circular SYN 20-AUG-2025
DEFINITION  Expresses H2A-Intein-Chitinbinding domain bound to H2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  TITLE     HisH2A-InteinCBD-H2B
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HisH2A-InteinCBD-H2B
FEATURES             Location/Qualifiers
     source          1..6923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             38..60
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             79..96
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             109..129
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             535..1128
                     /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             1159..1314
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     promoter        1388..1406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1407..1431
                     /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)."
     CDS             1474..1854
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             1868..1912
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1950..1968)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1964..2011
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2048..2503
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2135..2154)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2345..2366
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2618..3409,3410..3478))
                     /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     3241..3260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3479..3571)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3652..4240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4141..4160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4394..4411
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4426..4565
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4491..4510)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4651..4673
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4667..4858)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5430..6512)
                     /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     6474..6493
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6513..6590)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6796..6815
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6903..6921
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6922..6923,1..23)
                     /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)."
ORIGIN
        1 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
       61 tataccatgg gcagcagcca tcaccatcat caccacagcc aggatccaga gaacctctac
      121 ttccaaggca tgtcgggccg cggcaagact ggcggcaagg cccgcgccaa ggccaagtcg
      181 cgctcgtcgc gcgccggcct ccagttccca gtgggccgtg tacaccggct gctgcggaag
      241 ggccactacg ccgagcgcgt tggcgccggc gcgccagtgt acctggcggc agtgctggag
      301 tacctcaccg ctgagatcct ggagctggcg ggcaatgcgg cccgcgacaa caagaagacg
      361 cgaatcatcc cccgccacct gcagctggcc atccgcaacg acgaggagct caacaagctg
      421 ctgggcggcg tgacgatcgc ccagggaggc gtcctgccca acatccaggc cgtgctgctg
      481 cccaagaaga ccagcgccac cgtggggccg aaggcgccct cgggcggcaa gaagtgcatc
      541 acgggagatg cactagttgc cctacccgag ggcgagtcgg tacgcatcgc cgacatcgtg
      601 ccgggtgcgc ggcccaacag tgacaacgcc atcgacctga aagtccttga ccggcatggc
      661 aatcccgtgc tcgccgaccg gctgttccac tccggcgagc atccggtgta cacggtgcgt
      721 acggtcgaag gtctgcgtgt gacgggcacc gcgaaccacc cgttgttgtg tttggtcgac
      781 gtcgccgggg tgccgaccct gctgtggaag ctgatcgacg aaatcaagcc gggcgattac
      841 gcggtgattc aacgcagcgc attcagcgtc gactgtgcag gttttgcccg cgggaaaccc
      901 gaatttgcgc ccacaaccta cacagtcggc gtccctggac tggtgcgttt cttggaagca
      961 caccaccgag acccggacgc ccaagctatc gccgacgagc tgaccgacgg gcggttctac
     1021 tacgcgaaag tcgccagtgt caccgacgcc ggcgtgcagc cggtgtatag ccttcgtgtc
     1081 gacacggcag accacgcgtt tatcacgaac gggttcgtca gccacgctac tggcctcacc
     1141 ggtctgaact caggcctcac gacaaatcct ggtgtatccg cttggcaggt caacacagct
     1201 tatactgcgg gacaattggt cacatataac ggcaagacgt ataaatgttt gcagccccac
     1261 acctccttgg caggatggga accatccaac gttcctgcct tgtggcagct tcaatgaagc
     1321 ttgcggccgc ataatgctta agtcgaacag aaagtaatcg tattgtacac ggccgcataa
     1381 tcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc cccatcttag
     1441 tatattagtt aagtataaga aggagatata catatgccag agccagcgaa gtctgctccc
     1501 gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaaga cggcaagaag
     1561 cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc
     1621 caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac
     1681 attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc
     1741 atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag
     1801 cacgccgtgt ccgagggtac taaggccgtc accaagtaca ccagcgctaa gtagtctcga
     1861 gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag
     1921 cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1981 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     2041 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2101 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2161 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2221 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2281 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2341 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2401 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2461 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttctggcgg cacgatggca
     2521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3481 tcttcctttt tcaatcatga ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3541 atatttgaat gtatttagaa aaataaacaa ataggtcatg accaaaatcc cttaacgtga
     3601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3781 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4261 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4321 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4381 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4441 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     4501 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     4561 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4621 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4681 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4741 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4801 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4861 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4921 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4981 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     5041 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     5101 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     5161 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     5221 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     5281 ctagccgggt cctcaacgac aggagcacga tcatgctagt catgccccgc gcccaccgga
     5341 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5401 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5461 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5521 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5581 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5641 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5701 actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5761 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5821 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5881 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5941 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6001 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6061 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6121 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6181 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6241 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6301 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6361 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6421 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6481 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6541 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6601 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6661 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6721 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6781 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6841 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
     6901 attaatacga ctcactatag ggg
//
