LOCUS       sequence-407988-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Acidic Mammalian Chitinase catalytic domain with 
            C-terminal 6xHis tag in pcDNA 3.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  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 6563)
  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_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     859..878
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        859..877
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2092..2109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2150..2167)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2156..2380
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2426..2854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2513..2532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2723..2744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2863..2883)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2868..3197
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3048..3183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3110..3129
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3264..4058
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3318..3337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3928..3947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4232..4353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4269..4288)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4323..4342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4402..4418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4402..4418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4415..4437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4426..4442
                     /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)."
     promoter        complement(join(4450..4456,4457..4474,4475..4480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4495..4516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4633..4650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4804..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4884..4903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5563..6354,6355..6423))
                     /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     6186..6205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6424..6528)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
      121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      841 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt
      901 aaacttaagc ttggtaccat ggctaaactg ctgctggtca ctggtctggc actgctgctg
      961 aacgcacagc tggggtctgc ttacaacctg atttgctact tcaccaactg ggctcagtac
     1021 aggcctggcc tgggctcctt caagccagac gacatcaatc cttgcctgtg cacccacctg
     1081 atctacgcct ttgctggcat gcagaacaat gagatcacca caatcgagtg gaacgatgtg
     1141 accctgtata aggctttcaa tgacctgaag aacaggaact ccaagctgaa gacactgctg
     1201 gccatcggag gatggaactt cggcaccgct ccttttacca caatggtgag cacatctcag
     1261 aatagacaga cctttatcac aagcgtgatc aagttcctgc gccagtacgg ctttgatggc
     1321 ctggacctgg attgggagta tccaggcagc agaggctctc cccctcagga caagcatctg
     1381 ttcaccgtgc tggtgaagga gatgcgcgag gcctttgagc aggaggctat cgagtctaac
     1441 aggccaaggc tgatggtgac agctgctgtg gctggaggca tctctaatat ccaggctggc
     1501 tacgagatcc ccgagctgtc caagtatctg gatttcatcc acgtgatgac ctacgacctg
     1561 catggctcct gggagggcta tacaggcgag aacagcccac tgtacaagta tcccaccgag
     1621 acaggctcca acgcctacct gaatgtggat tacgtgatga attattggaa gaacaatggc
     1681 gcccctgctg agaagctgat cgtgggcttc cctgagtatg gccacacctt tatcctgaga
     1741 aacccttctg ataatggcat cggcgcccca acctccggcg acggacctgc tggaccatac
     1801 acaaggcagg ccggcttctg ggcttactat gagatttgca cctttctgcg gagcggagct
     1861 acagaagtgt gggatgcttc tcaggaggtg ccatacgcct ataaggctaa cgagtggctg
     1921 ggctatgaca atatcaagtc cttcagcgtg aaggctcagt ggctgaagca gaacaatttt
     1981 ggcggcgcca tgatctgggc tatcgatctg gacgatttca ccggcagctt ttgtgaccag
     2041 ggcaagttcc ccctgacctc cacactgaac aaggccctgg gcatcagcac ccatcaccat
     2101 catcaccatt gatctagatc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2161 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2221 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2281 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2341 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2401 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2641 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2701 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2761 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2821 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3001 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3061 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3121 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3181 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3241 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3301 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3361 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3421 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3481 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3541 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3601 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3661 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3721 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3781 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3841 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3901 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3961 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4021 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4081 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4141 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4201 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4261 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4321 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4381 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6541 aaaagtgcca cctgacgtcg acg
//
