LOCUS       sequence-407989-        5391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Acidic Mammalian Chitinase catalytic domain with 
            C-terminal 6xHis tag in pTwist CMV vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  AUTHORS   Diaz RE, Ecker AK, Correy GJ, Asthana P, Young ID, Faust B, Thompson
            MC, Seiple IB, Van Dyken S, Locksley RM, Fraser JS
  TITLE     Structural characterization of ligand binding and pH-specific 
            enzymatic activity of mouse Acidic Mammalian Chitinase.
  JOURNAL   Elife. 2024 Jun 17;12:RP89918. doi: 10.7554/eLife.89918.
  PUBMED    38884443
REFERENCE   2  (bases 1 to 5391)
  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: "Elife."; 
            date: "2024-06-17"; volume: "12:RP89918. doi"; pages: " 
            10.7554/eLife.89918"
COMMENT     Sequence Label: mAMCase_CatD_6xHis_pTwistCMV
FEATURES             Location/Qualifiers
     source          1..5391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        111..313
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     264..284
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1557..1574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1643..1764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1654..1673)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1708..1727
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1879..1925
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1955..2091
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2136..2207
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2433..3021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2513..2532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3767..3835,3836..4627)
                     /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     complement(3985..4004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4640..4776
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4928..4999
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5004..5050)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5068..5116
                     /note="synthetic polyadenylation signal"
     enhancer        join(5197..5391,1..109)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
       61 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
      121 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      181 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      241 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      301 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
      361 tcgacacacc cgccagcggc cgcatggcaa agctcctgct tgtcactgga ctcgccctgc
      421 tgctgaacgc tcagctcggg tctgcttaca acctgatatg ctatttcact aactgggcgc
      481 agtatagacc cggtctgggt agcttcaagc ctgatgatat taacccatgt ctttgtaccc
      541 acctgatcta cgcgtttgcc ggcatgcaga ataatgagat aaccaccatt gagtggaatg
      601 atgtaacact gtataaggcc ttcaatgacc ttaagaaccg caacagcaag ctgaaaacat
      661 tactcgccat aggcggatgg aactttggta cagccccctt cacaaccatg gtttctactt
      721 cccagaacag acaaacattt atcacctctg taatcaaatt tctccggcag tatggcttcg
      781 acgggctgga cttagattgg gagtatccag gttccagggg atccccccct caagataagc
      841 acttgttcac tgtcctggtg aaggagatga gagaggcatt tgagcaggaa gctattgaat
      901 ctaataggcc ccgactgatg gtgaccgccg cagtggccgg cggaatctca aacatccaag
      961 ccggctatga gatcccagaa ctgtccaaat atctcgattt tattcatgtg atgacctatg
     1021 atcttcatgg ctcttgggaa ggatatacag gagaaaattc tcccctgtat aaatatccaa
     1081 ccgaaactgg ttctaacgcc tacctcaatg tggattacgt catgaactac tggaagaata
     1141 acggggctcc tgctgagaaa ttaattgtag gttttccaga atacgggcac accttcatct
     1201 tacggaaccc aagtgataat ggaattggcg cccccacgtc tggggatggt cccgccggac
     1261 catacactag acaagcgggt ttttgggcgt actacgagat ttgtacattt ctgagatctg
     1321 gggccactga ggtatgggat gcaagtcagg aagtcccgta cgcctataag gccaatgaat
     1381 ggctcgggta tgacaacatc aagtcctttt cagtgaaggc tcaatggctt aaacagaaca
     1441 atttcggcgg tgcgatgatt tgggcaatcg acctggacga ctttacaggg tctttctgtg
     1501 accaggggaa gtttccactg acgagcaccc tgaataaggc gctcggaatt agcacacacc
     1561 accatcacca ccattgagga tccgcaggcc tctgctagct tgactgactg agatacagcg
     1621 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1681 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1741 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1801 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     1861 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1921 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     1981 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     2041 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     2101 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     2161 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     2221 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     2281 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     2341 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     2401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3181 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3241 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     3301 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     3361 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     3421 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     3481 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     3541 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     3601 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     3661 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     3721 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     3781 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3841 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     3901 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     3961 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4021 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4081 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4141 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     4201 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4261 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4321 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     4381 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4441 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4501 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4561 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4621 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     4681 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     4741 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     4801 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     4861 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     4921 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     4981 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5041 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     5101 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     5161 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5221 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5281 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5341 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta t
//
