LOCUS       sequence-484470-        4955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse IgG2a heavy chain constant region. Users can clone 
            in their desired heavy chain variable region for mammalian 
            expression; to be used with pAG mKappa Empty light chain (Plasmid 
            242954).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4955)
  AUTHORS   Rego M, Fan M, Addgene Research Program
  TITLE     Addgene Antibody Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4955)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAG mIgG2a Empty
FEATURES             Location/Qualifiers
     source          1..4955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     enhancer        155..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        535..738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     689..709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          840..972
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             1395..2090
                     /codon_start=1
                     /product="CH2 and CH3 domains of the mouse IgG2a heavy 
                     chain plus the hinge region"
                     /label=Fc tag
                     /translation="PRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVT
                     CVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEF
                     KCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIY
                     VEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHT
                     TKSFSRTPGK"
     misc_feature    2124..2712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2177..2197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2595..2606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2740..2759)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2792..2840
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(2965..2982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3136..3724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3216..3235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3895..4686,4687..4755))
                     /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     4518..4537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4756..4860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4928..4946
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactaac cggtgacatt gattattgac tagttattaa
      181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      721 gtctatataa gcagagctcc taggcgttta gtgaaccgtc agatcgcctg gagacgccat
      781 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag
      841 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
      901 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
      961 ttctctccac agggcgcgcc gtgtccaggg ccaccgaatt catgacccgg ctgacagtcc
     1021 tggccctgct ggctggtctg ctggcgtcct cgagggccat ccgatcctta gcggttcact
     1081 gatcgaacgt ggaagtctac gcgcgcccaa cagccccatc ggtctatcca ctggcccctg
     1141 tgtgtggaga tacaactggc tcctcggtga ctctaggatg cctggtcaag ggttatttcc
     1201 ctgagccagt gaccttgacc tggaactctg gatccctgtc cagtggtgtg cacaccttcc
     1261 cagctgtcct gcagtctgac ctctacaccc tcagcagctc agtgactgta acctcgagca
     1321 cctggcccag ccagtccatc acctgcaatg tggcccaccc ggcaagcagc accaaggtgg
     1381 acaagaaaat tgagcccaga gggcccacaa tcaagccctg tcctccatgc aaatgcccag
     1441 cacctaacct cttgggtgga ccatccgtct tcatcttccc tccaaagatc aaggatgtac
     1501 tcatgatctc cctgagcccc atagtcacat gtgtggtggt ggatgtgagc gaggatgacc
     1561 cagatgtcca gatcagctgg tttgtgaaca acgtggaagt acacacagct cagacacaaa
     1621 cccatagaga ggattacaac agtactctcc gggtggtcag tgccctcccc atccagcacc
     1681 aggactggat gagtggcaag gagttcaaat gcaaggtcaa caacaaagac ctcccagcgc
     1741 ccatcgagag aaccatctca aaacccaaag ggtcagtaag agctccacag gtatatgtct
     1801 tgcctccacc agaagaagag atgactaaga aacaggtcac tctgacctgc atggtcacag
     1861 acttcatgcc tgaagacatt tacgtggagt ggaccaacaa cgggaaaaca gagctaaact
     1921 acaagaacac tgaaccagtc ctggactctg atggttctta cttcatgtac agcaagctga
     1981 gagtggaaaa gaagaactgg gtggaaagaa atagctactc ctgttcagtg gtccacgagg
     2041 gtctgcacaa tcaccacacg actaagagct tctcccggac tccgggtaaa tgaaagcttt
     2101 gagtttgata tctcgacggt accaatcaac ctctggatta caaaatttgt gaaagattga
     2161 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2221 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2281 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2341 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2401 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2461 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     2521 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2581 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2641 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2701 ccgcctcccc gcacgcgtct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     2761 accggaagga acccgcgcta tgagcggccg ccggcaataa aaagacagaa taaaacgcac
     2821 gggtgttggg tcgtttgttc ttaattaaat tgcgttgcgc tcactgcccg ctttccagtc
     2881 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2941 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3001 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3061 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3541 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4861 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4921 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
