LOCUS       sequence-392892-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing an antigen targeting the GABA(B)R2 metabotropic 
            GABA receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  TITLE     Trimmer lab antigen plasmids
REFERENCE   2  (bases 1 to 8098)
  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"
COMMENT     Sequence Label: GABA BR2-flag/pRK5
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /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)."
     primer_bind     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        431..810
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        811..1014
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1011..1035
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1245..1263
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1245..1262
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1276..1295
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1847..1870
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4815..4824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4828..4962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4865..4884)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4919..4938
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5031..5388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5031..5222
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5054..5074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5239..5374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5408..5425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5408..5424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5417..5439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5637..6092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5724..5743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5934..5955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6109..6128)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6228..6250
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6288..6306)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6374..6478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6479..6547,6548..7339)
                     /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     complement(6697..6716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7510..8098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7999..8018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc
      421 gagctcgccc gacattgatt attgtctaga tattaatagt aatcaattac ggggtcatta
      481 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      541 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      601 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      661 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      721 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      781 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      841 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      901 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      961 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     1021 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1081 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
     1141 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa
     1201 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     1261 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc
     1321 tcggttctat cgattgaatt ccccggggat cctctagagt cgacccgggc ggccgcgctg
     1381 gagagaggcg cgcggagact cggccccctc ggcgctcgct cgctcgctgg ctccccgcct
     1441 cccgcactcc gctggctccc accccttccc ggcgtgattg atccgtcacg ggcgccgcca
     1501 ccgccgcggc cgttctgagc gagcagaagc cggagccgag ccggggcccg tgccggcgcc
     1561 attgcgcggg agccgcgggc aaagctcggc gccgggcggc gggccgggcc aggccatgcg
     1621 ggccgagtga gctggcgccc gcagcccgcg gcgcggcatg gcttccccgc cgagctccgg
     1681 gcagccccgg ccgccgccgc cgccgccgcc gccgcccgcg cgcctgctgc tgcccctgct
     1741 gctgtcgctg ctgctgtggt tggcgcccgg ggcctggggc tggacgcggg gcgccccccg
     1801 gccgccgccc agcagcccgc cgctctccat catgggcctc atggtcgact acaaggacga
     1861 cgatgacaag ccgctcacca aggaggtggc caagggcagc atcgggcgcg gcgtgctccc
     1921 cgccgtggag ctagccatcg agcagatccg caacgagtca ctcctgcgcc cctacttcct
     1981 ggacctgcga ctctatgaca ccgagtgtga caatgcaaag ggactgaaag ccttctatga
     2041 cgcaataaag tatgggccga accatttgat ggtgtttgga ggcgtctgtc cgtctgtcac
     2101 atctattatc gcggagtccc tccaaggctg gaatctggtg cagctttcct tcgccgccac
     2161 cacgcctgtt cttgcggata agaagaagta cccgtatttc ttccggacgg tgccgtcaga
     2221 caacgcggtg aaccccgcca tcctgaagct cctgaagcac ttccgctggc ggcgtgtggg
     2281 cacactcacg caggacgtgc agcgcttctc cgaggtgagg aatgacctga ctggggttct
     2341 gtatggggaa gatattgaga tctcagacac agagagcttc tccaatgatc cctgcaccag
     2401 cgtcaaaaag ctcaagggga atgacgtgcg gatcatcctt ggccagtttg accagaatat
     2461 ggcagcaaaa gtcttctgtt gtgccttcga ggagagcatg tttggcagca agtaccagtg
     2521 gatcatcccg ggatggtacg agcctgcgtg gtgggagcag gtgcatgtgg aggccaattc
     2581 ctcacgctgc ctgcgcagaa gcctcctggc tgccatggaa ggttacatcg gagtggactt
     2641 tgagcccctg agctccaaac aaatcaagac catctcaggg aagactccac agcagtatga
     2701 aagagagtac aacaccaaac gttcaggcgt ggggcccagc aagttccatg ggtacgccta
     2761 cgatgggatc tgggtcatcg ccaagaccct acagagggcc atggagacac tgcatgccag
     2821 tagcaggcac cagcggatcc aggacttcaa ctacacagac cacacgctgg gcaaaatcat
     2881 cctcaatgcc atgaacgaga ccaacttctt cggggtcacg ggtcaagttg tgttccggaa
     2941 cggggagaga atgggaacca ttaaatttac tcaatttcaa gacagcagag aggtgaaggt
     3001 cggcgaatac aacgcggtgg ctgacacact ggagatcatc aatgacacca taaggttcca
     3061 ggggtccgag ccacccaagg acaagaccat cattctggag cagcttcgga agatctcgct
     3121 tccactgtat agcatcctgt ccgctctcac catcctcggc atgatcatgg ccagcgcctt
     3181 cctcttcttc aacatcaaga accggaacca aaagctgatt aagatgtcaa gcccctacat
     3241 gaacaacctc atcatcctgg gaggaatgct gtcctatgca tccatcttcc tctttggcct
     3301 cgatgggtcc ttcgtctcag aaaagacctt tgaaacactc tgcacggtcc ggacctggat
     3361 tctcaccgtg ggctacacaa ctgcctttgg ggccatgttt gcaaagacct ggagggtcca
     3421 tgccatcttc aaaaatgtga agatgaagaa gaagatcatc aaagaccaga agctgcttgt
     3481 gattgtgggg ggcatgctgc tcatcgacct gtgcatcctg atctgttggc aggctgtgga
     3541 ccccctgcgg aggacagtag agaggtacag catggagccg gacccagcag gccgggacat
     3601 ctccatccgc ccattgctgg aacactgcga aaacacccac atgaccatct ggcttggcat
     3661 tgtctacgcc tacaaggggc tcctcatgct attcggttgt ttcttggcat gggaaacccg
     3721 caatgtgagc atccctgccc tcaacgacag caagtacatc ggcatgagtg tgtacaatgt
     3781 ggggatcatg tgcatcatcg gggctgctgt ctccttcctg acgcgtgacc agcccaacgt
     3841 gcagttctgc atcgtggccc tggtcatcat cttctgcagc accatcactc tctgcctggt
     3901 gtttgtgcca aagctcatca ctctgaggac aaaccctgac gcagccactc agaacaggcg
     3961 gttccagttc acacagaacc agaagaaaga agattcgaag acctccactt cagtcaccag
     4021 cgtgaaccag gcgagcacgt cacgcctgga gggactgcag tcagaaaacc accgccttcg
     4081 aatgaagatc acagagctgg acaaagactt ggaagaagtc accatgcagc tacaagacac
     4141 accagagaag accacataca tcaaacagaa tcactaccaa gagctcaacg acatcctcag
     4201 cttgggcaac ttcacagaga gcacagatgg aggaaaggcc attctaaaaa atcacctcga
     4261 tcaaaacccc cagctccagt ggaacacgac agagccctca agaacatgca aagaccccat
     4321 agaagacatc aactccccgg agcacatcca gcgccggctg tcgctccagc tccccatcct
     4381 tcaccacgcc tacctcccat ccatcggagg cgtggatgcc agctgcgtca gcccctgtgt
     4441 cagccctacc gccagccctc gccacagaca cgtaccaccc tccttccgag tcatggtctc
     4501 gggcctgtag gggtgggagg cctggggccg gggcctcccc gggacagcac catgctgggc
     4561 caaggcgcct gccacaggca cactgacggc ggcgagaagc tgggcaccat gctgcctctc
     4621 cagactgctg gaatggcgct caggcagagc gggactcggc accgacctcg agccttatct
     4681 gtgaaggtct tactctcaca gaggagagga atgacaatga cttctccttc ttggcgtctg
     4741 caaacaaaga ggagttggga tgtctgaaac ttgcaaaaac aaatcaaact ctagagtcga
     4801 cctgcagaag cttggccgcc atggcccaac ttgtttattg cagcttataa tggttacaaa
     4861 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4921 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcgggaatta attcggcgca
     4981 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     5041 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctgttaaca
     5401 gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5461 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5521 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     5581 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     5641 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5701 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5761 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5821 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5881 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5941 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     6001 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6061 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     6121 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     6181 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6241 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     6301 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6721 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7021 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgctacgct
     7921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaa
//
