LOCUS       sequence-370115-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MUC2 D1D2D' assembly with cleavable His tag in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  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 8069)
  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-MUC2 D1D2D' with cleavable His tag
FEATURES             Location/Qualifiers
     source          1..8069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        872..890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1043..1060
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1061..1081
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3669..3686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3675..3899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3945..4373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4032..4051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4242..4263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4382..4402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4387..4716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4567..4702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4629..4648
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4783..5577
                     /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(4837..4856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5447..5466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5751..5872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5788..5807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5842..5861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5921..5937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5921..5937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5934..5956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5945..5961
                     /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(5969..5975,5976..5993,5994..5999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6014..6035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6152..6169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6323..6911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6403..6422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7082..7873,7874..7942))
                     /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     7705..7724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7943..8047)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      841 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      901 ctggctagcg tttaaactta agcttggtac cgagctcgga tccatgagga ggtgcaacag
      961 cggctccggg ccgccgccgt cgctgctgct gctgttgctg tggctgctcg cggttcccgg
     1021 cgctaacgcg gccccgcagg gtcatcatca tcaccatcac gagaatctct atttccaggg
     1081 atcggagctc cagacagagg gcagaacccg aaaccacggc cacaacgtct gcagcacctg
     1141 gggcaacttc cactacaaga ccttcgacgg ggacgtcttc cgcttccccg gcctctgcga
     1201 ctacaacttc gcctccgact gccgaggctc ctacaaggaa tttgctgtgc acctgaagcg
     1261 gggtccgggc caggctgagg cccccgccgg ggtggagtcc atcctgctga ccatcaagga
     1321 tgacaccatc tacctcaccc gccacctggc tgtgcttaac ggggccgtgg tcagcacccc
     1381 gcactacagc cccgggctgc tcattgagaa gagcgatgcc tacaccaaag tctactcccg
     1441 cgccggcctc accctcatgt ggaaccggga ggatgcactc atgctggagc tggacactaa
     1501 gttccggaac cacacctgtg gcctctgcgg ggactacaac ggcctgcaga gctattcaga
     1561 attcctctct gacggcgtgc tcttcagtcc cctggagttt gggaacatgc agaagatcaa
     1621 ccaacccgat gtggtgtgtg aggatcccga ggaggaggtg gcccccgcat cctgctccga
     1681 gcaccgcgcc gagtgtgaga ggctgctgac cgccgaggcc ttcgcggact gtcaggacct
     1741 ggtgccgctg gagccgtatc tgcgcgcctg ccagcaggac cgctgccggt gcccgggcgg
     1801 tgacacctgc gtctgcagca ccgtggccga gttctcccgc cagtgctccc acgccggcgg
     1861 ccggcccggg aactggagga ccgccacgct ctgccccaag acctgccccg ggaacctggt
     1921 gtacctggag agcggctcgc cctgcatgga cacctgctca cacctggagg tgagcagcct
     1981 gtgcgaggag caccgcatgg acggctgttt ctgcccagaa ggcaccgtat atgacgacat
     2041 cggggacagt ggctgcgttc ctgtgagcca gtgccactgc aggctgcacg gacacctgta
     2101 cacaccgggc caggagatca ccaatgactg cgagcagtgt gtctgtaacg ctggccgctg
     2161 ggtgtgcaaa gacctgccct gccccggcac ctgtgccctg gaaggcggct cccacatcac
     2221 caccttcgat gggaagacgt acaccttcca cggggactgc tactatgtcc tggccaaggg
     2281 tgaccacaac gattcctatg ctctcctggg cgagctggcc ccctgtggct ccacagacaa
     2341 gcagacctgc ctgaagacgg tggtgctgct ggctgacaag aagaagaatg tggtggtctt
     2401 caagtccgat ggcagtgtac tgctcaacga gctgcaggtg aacctgcccc acgtgaccgc
     2461 gagcttctct gtcttccgcc cgtcttccta ccacatcatg gtgagcatgg ccattggcgt
     2521 ccggctgcag gtgcagctgg ccccagtcat gcaactcttt gtgacactgg accaggcctc
     2581 ccaggggcag gtgcagggcc tctgcgggaa cttcaacggc ctggaaggtg acgacttcaa
     2641 gacggccagc gggctggtgg aggccacggg ggccggcttt gccaacacct ggaaggcaca
     2701 gtcaagctgc catgacaagc tggactggtt ggacgatccc tgctccctga acatcgagag
     2761 cgccaactac gccgagcact ggtgctccct cctgaagaag acagagaccc cctttggcag
     2821 gtgccactcg gctgtggacc ctgctgagta ttacaagagg tgcaaatatg acacgtgtaa
     2881 ctgtcagaac aatgaggact gcctgtgcgc cgccctgtcc tcctacgcgc gcgcctgcac
     2941 cgccaagggc gtcatgctgt ggggctggcg ggagcatgtc tgcaacaagg atgtgggctc
     3001 ctgccccaac tcgcaggtct tcctgtacaa cctgaccacc tgccagcaga cctgccgctc
     3061 cctctccgag gccgacagcc actgtctcga gggctttgcg cctgtggacg gctgcggctg
     3121 ccctgaccac accttcctgg acgagaaggg ccgctgcgta cccctggcca agtgctcctg
     3181 ttaccaccgc ggtctctacc tggaggcggg ggatgtggtc gtcaggcagg aagaacgatg
     3241 tgtgtgccgg gatgggaggc tgcactgtag gcagatccgg ctgatcggcc agagctgcac
     3301 ggccccaaag atccacatgg actgcagcaa cctgactgca ctggccacct cgaagccccg
     3361 agccctcagc tgccagacgc tggccgccgg ctattaccac acagagtgtg tcagtggctg
     3421 tgtgtgcccc gacgggctga tggatgacgg ccggggtggc tgtgtggtgg agaaggaatg
     3481 cccttgcgtc cataacaacg acctgtattc ttccggcgcc aagatcaagg tggactgcaa
     3541 tacctgcacc tgcaagagag gacgctgggt gtgcacccag gctgtgtgcc atggctgaga
     3601 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     3661 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3721 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3781 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3841 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3901 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5881 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6721 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6841 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6901 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6961 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7021 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7081 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7141 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7201 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7261 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7321 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7381 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7441 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7501 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7561 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7621 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7681 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7741 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7801 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7861 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7921 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7981 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8041 ttccgcgcac atttccccga aaagtgcca
//
