LOCUS       sequence-211628-        7523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human LRRC33 in 293T cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7523)
  AUTHORS   Qin Y, Garrison BS, Ma W, Wang R, Jiang A, Li J, Mistry M, Bronson 
            RT, Santoro D, Franco C, Robinton DA, Stevens B, Rossi DJ, Lu C, 
            Springer TA
  TITLE     A Milieu Molecule for TGF-beta Required for Microglia Function in 
            the Nervous System.
  JOURNAL   Cell. 2018 Jun 28;174(1):156-171.e16. doi: 
            10.1016/j.cell.2018.05.027. Epub 2018 Jun 14.
  PUBMED    29909984
REFERENCE   2  (bases 1 to 7523)
  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.2018.05.027"; journalName: "Cell"; date: 
            "2018-06-28- 28"; volume: "174"; issue: "1"; pages: "156-171.e16"
COMMENT     Sequence Label: pcDNA3.1(+)-human LRRC33
FEATURES             Location/Qualifiers
     source          1..7523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        164..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        544..747
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     698..718
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     792..811
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        792..810
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2956..2985
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3001..3018
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3041..3058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3047..3271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3317..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3614..3635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3754..3774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3759..4088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3939..4074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4001..4020
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4155..4949
                     /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(4209..4228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4819..4838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5125..5246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5162..5181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5216..5235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5295..5311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5295..5311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5308..5330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5319..5335
                     /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(5343..5349,5350..5367,5368..5373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5388..5409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5526..5543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5697..6285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5777..5796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6456..7247,7248..7316))
                     /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     7079..7098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7317..7421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7496..7515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
       61 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
      121 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      721 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
      781 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagtt aagcttggta
      841 ccgagctcgg atcctgccct tgagatggag ttgctgcctc tttggctctg cctgggtttt
      901 cacttcctga ccgtgggctg gaggaacaga agcggaacag ccacagcagc ctcccaagga
      961 gtctgcaagt tggtgggtgg agccgctgac tgccgagggc agagcctcgc ttcggtgccc
     1021 agcagcctcc cgccccacgc ccggatgctc accctggatg ccaaccctct caagaccctg
     1081 tggaatcact ccctccagcc ttaccctctc ctggagagcc tcagcctgca cagctgccac
     1141 ctggagcgca tcagccgcgg cgccttccag gagcaaggtc acctgcgcag cctggtactg
     1201 ggggacaact gcctctcaga gaactacgaa gagacggcag ccgccctcca cgccctgccg
     1261 ggcctgcgga ggctggactt gtcaggaaac gccctgacgg aggacatggc agcgctcatg
     1321 ctccagaacc tctcctcgct gcggtccgtg tccctggcgg ggaacaccat catgcggctg
     1381 gacgactccg tcttcgaggg cctggagcgt ctccgggagc tggatctgca gaggaactac
     1441 atcttcgaga tcgagggcgg cgctttcgac ggcctggctg agctgaggca cctcaacctg
     1501 gccttcaaca acctcccctg catcgtggac ttcgggctca cgcggctgcg ggtcctcaac
     1561 gtcagctaca acgtcctgga gtggttcctc gcgaccgggg gagaggctgc cttcgagctg
     1621 gagacgctgg acctgtctca caaccagctg ctgttcttcc cgctgctgcc ccagtacagc
     1681 aagttgcgga ccctcctgct gcgcgacaac aacatgggct tctaccggga cctgtacaac
     1741 acctcgtcgc cgagggagat ggtggcccag ttcctcctcg tggacggcaa cgtgaccaac
     1801 atcaccaccg tcagcctctg ggaagaattc tcctccagcg acctcgcaga tctccgcttc
     1861 ctggacatga gccagaacca gttccagtac ctgccagacg gcttcctgag gaaaatgcct
     1921 tccctctccc acctgaacct ccaccagaat tgcctgatga cgcttcacat tcgggagcac
     1981 gagccccccg gagcgctcac cgagctggac ctgagccaca accagctgtc ggagctgcac
     2041 ctggctccgg ggctggccag ctgcctgggc agcctgcgct tgttcaacct gagctccaac
     2101 cagctcctgg gcgtcccccc tggcctcttc gccaatgcta ggaacatcac tacacttgac
     2161 atgagccaca atcagatctc actttgtccc ctgccagctg cctcggaccg ggtgggcccc
     2221 cctagctgtg tggatttcag gaatatggca tctttaagga gcctgtctct ggagggctgt
     2281 ggcctggggg cattgccaga ctgcccattc caagggacct ccctgaccta cttagacctc
     2341 tcaagcaact ggggggttct gaatgggagc ctcgccccac tccaggatgt tgcccccatg
     2401 ttacaggtcc tgtctctcag gaacatgggc ctccactcca gctttatggc gttggacttc
     2461 tctgggtttg ggaatctcag ggacttagat ctgtcgggga attgcttgac caccttccca
     2521 aggtttgggg gcagcctggc cctggagacc ctggatctcc gtagaaactc gctcacagcc
     2581 cttccccaga aggctgtgtc tgagcagctc tcgagaggtc tgcggaccat ctacctcagt
     2641 cagaatccat atgactgctg tggggtggat ggctgggggg ccctgcagca tgggcagacg
     2701 gtggccgact gggccatggt cacctgcaac ctctcctcca agatcatccg cgtgacggag
     2761 ctgcccggag gtgtgcctcg ggactgcaag tgggagcggc tggacctggg cctgctctac
     2821 ctcgtgctca tcctccccag ctgcctcacc ctgctggtgg cctgcactgt catcgtcctc
     2881 acttttaaga agcctctgct tcaggtcatc aagagccgct gccactggtc ctccgtttac
     2941 tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat gcataccggt
     3001 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     3061 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3121 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3181 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3241 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3301 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3361 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3421 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3481 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3541 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3601 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3661 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3721 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4021 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4081 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4141 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4201 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4261 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4321 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4381 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4441 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4501 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4561 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4621 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4681 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4741 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4801 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4861 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4921 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4981 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5041 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5101 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5161 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5221 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5281 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5341 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5401 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5461 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5521 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5581 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6361 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6421 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6481 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6541 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6601 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6661 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6721 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6781 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6841 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6901 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6961 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7021 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7081 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7141 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7201 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7261 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7321 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7381 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7441 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7501 tctgctctga tgccgcatag tta
//
