LOCUS       sequence-355108-        6680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Muc5b D1 with cleavable His tag, residues 50-426.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6680)
  AUTHORS   Reznik N, Gallo AD, Rush KW, Javitt G, Fridmann-Sirkis Y, Ilani T, 
            Nairner NA, Fishilevich S, Gokhman D, Chacon KN, Franz KJ, Fass D
  TITLE     Intestinal mucin is a chaperone of multivalent copper.
  JOURNAL   Cell. 2022 Oct 27;185(22):4206-4215.e11. doi: 
            10.1016/j.cell.2022.09.021. Epub 2022 Oct 6.
  PUBMED    36206754
REFERENCE   2  (bases 1 to 6680)
  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: 
            "10.1016/j.cell.2022.09.021"; journalName: "Cell"; date: 
            "2022-10-27- 27"; volume: "185"; issue: "22"; pages: "4206-4215.e11"
COMMENT     Sequence Label: pcDNA3.1-Muc5b D1 (murine) with cleavable His tag
FEATURES             Location/Qualifiers
     source          1..6680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        179..558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        559..762
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     713..733
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     807..826
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        807..825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2103..2123
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2124..2141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2215..2232)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2221..2445
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2491..2919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2578..2597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2788..2809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2928..2948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2933..3262
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3113..3248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3175..3194
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3329..4123
                     /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(3383..3402)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3993..4012
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4297..4418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4334..4353)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4388..4407
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4467..4483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4467..4483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4480..4502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4491..4507
                     /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(4515..4521,4522..4539,4540..4545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4560..4581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4698..4715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4869..5457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4949..4968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5628..6419,6420..6488))
                     /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     6251..6270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6489..6593)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(6668..6680,1..7))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cactgagtag
       61 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      121 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      181 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      241 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      301 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      361 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      421 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      481 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      541 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      601 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      661 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      721 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      781 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
      841 tagcgtttaa acttaagctt ggtaccgagc tcggatccat gaggaggtgc aacagcggct
      901 ccgggccgcc gccgtcgctg ctgctgctgt tgctgtggct gctcgcggtt cccggcgcta
      961 acgcggcccc gatatcctca tccacaaggc atgtcacctt catcccacct atcactgtct
     1021 tccctagcat gagcgcctta cacctggcac acaatgggag agtgtgcagc acgtggggtg
     1081 acttccatta caagaccttt gatggtgacg tgttccgctt ccctggcctc tgtaactatg
     1141 tcttctcttc acactgcggg gccacctacg aggacttcaa catccaactg cgtcgtggtc
     1201 ttgagggttc caggcccaca gtcacctatg tgctcctcag ggcccagggg ctggtgattg
     1261 agctgtccaa tggctctgtc ctggtgaatg gacatcggga gaagctgccc tacagtcgtg
     1321 caggtctcct gatggagaag agcagtggct atgtgaaaat cagcatccgc ctagtcctca
     1381 ccttcctgtg gaatgaagaa gacagcgccc tgctggagct ggactccaaa tacatcaacc
     1441 agacgtgtgg gctgtgcggt gacttcaacg ggctcccagc tgtcagtgag ttctacaccc
     1501 acaacactag gctgacccct gtgcagtttg gaaacttgca gaagctggat ggtcctacag
     1561 agcagtgcca ggatgtccta ccctcagctg tcagcaactg cacagatacg gaggacatct
     1621 gccgtcgcac cctgcttggt cctgcctttg acaagtgcac cgctctggta gatgtcagca
     1681 tgtacttgga tgcctgtgtc caggacctgt gccgctgtcc cacctgccca tgtgccactt
     1741 ttgccgaata ttcccgccag tgtgcccacg cagggggcta cccacagaac tggaggcatc
     1801 ctgatctctg caattggaca tgccctttca acatggaata ccgagagtgc agttcaccct
     1861 gtgtggatac ctgctctaac ccccagcgct ctcaactctg cgaggaccac tgcatggatg
     1921 gctgcttctg tcccccagga actgtgctgg acgatgtcag acacttggga tgcctgcccc
     1981 tggagcagtg ccactgtaca cacggtggcc gaatttatgc ccctggagag tccttcaaca
     2041 ccagctgtag gtcctgcacc tgcttcggag gactgtggaa gtgcaaagac ctgccatgcc
     2101 ctgagaatct ctatttccag ggacatcatc atcaccatca ctgagaattc tgcagatatc
     2161 cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2221 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2281 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2341 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2401 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2461 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2521 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2581 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2641 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2701 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2761 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2821 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2881 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2941 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3001 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3121 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3181 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3241 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3301 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3361 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3421 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3481 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3541 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3601 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3661 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3721 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3781 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3841 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3901 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3961 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4021 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4081 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4141 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4201 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4261 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4321 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4381 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4441 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4501 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4561 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4621 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4681 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4741 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4801 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4861 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4921 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4981 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5041 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5101 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5161 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5221 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5281 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5341 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5401 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5461 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5521 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5581 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5641 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5701 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5761 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5821 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5881 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5941 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6001 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6061 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6121 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6181 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6241 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6301 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6361 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6421 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6481 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6541 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6601 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6661 ctctcagtac aatctgctct
//
