LOCUS       sequence-309787-        7224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HA-3xFLAG-VFT-TMD-CC in pFastBac1 for Sf9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7224)
  AUTHORS   Shaye H, Ishchenko A, Lam JH, Han GW, Xue L, Rondard P, Pin JP, 
            Katritch V, Gati C, Cherezov V
  TITLE     Structural basis of the activation of a metabotropic GABA receptor.
  JOURNAL   Nature. 2020 Jun 17. pii: 10.1038/s41586-020-2408-4. doi: 
            10.1038/s41586-020-2408-4.
  PUBMED    32555460
REFERENCE   2  (bases 1 to 7224)
  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: "Nature. 
            2020 Jun 17. pii: 10.1038/s41586-020-2408-4. doi: 
            10.1038/s41586-020-2408-4."
COMMENT     Sequence Label: IMPT-10314:GABBR2
FEATURES             Location/Qualifiers
     source          1..7224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  1..225
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(292..825)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(995..1000,1001..1017,1018..1023))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1375..1466
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1391..1408
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1619..1684
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1688..1708
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4204..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4241..4260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4295..4314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4367..4532
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4716..5170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4803..4822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5012..5033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5196..5300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5301..5369,5370..6161)
                     /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(5519..5538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6332..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7074..7091
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
       61 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
      121 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
      181 gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa
      241 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
      301 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
      361 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
      421 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
      481 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
      541 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
      601 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
      661 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
      721 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
      781 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
      841 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata
      901 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga
      961 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt
     1021 caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag
     1081 cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac
     1141 gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct
     1201 gccctggctt caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac
     1261 cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca
     1321 ggactctagc tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg
     1381 gagataatta aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca
     1441 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc
     1501 gcggatcccg gtccgaagcg cgcggaattc aaaggcctac gtcgacgagc tcactagtcg
     1561 cggccgccac atgaaaacaa ttatagctct cagctacata ttttgtctgg tgttcgcgga
     1621 ttataaggac cacgatggag actataaaga ccatgacatt gattataagg atgatgacga
     1681 taaattggag aacttgtact tccagggatg ggctcgcggt gcaccccgtc ctcccccatc
     1741 cagcccgcct ctgtctatca tgggattgat gccactgacc aaggaggtgg ctaaaggttc
     1801 aatcggaaga ggagttctgc cagcagtgga gctcgcaatc gaacagattc gcaacgagtc
     1861 cctgctccgt ccctacttct tggacctgcg tctctacgac accgaatgcg ataacgccaa
     1921 gggcctcaaa gcgttctacg atgctatcaa gtacggaccc aaccacttga tggtcttcgg
     1981 tggcgtttgt ccaagtgtga cttcgatcat tgctgagagt ctccagggct ggaacctcgt
     2041 gcaattgtcg ttcgctgcca ccactccagt cttggccgac aagaaaaagt acccgtactt
     2101 cttccgcacc gtgccttccg ataacgccgt caaccccgca atcctgaagt tgctgaaaca
     2161 ttaccagtgg aagcgtgtcg gaaccctcac tcaggacgtt caacgcttca gcgaggtgcg
     2221 taacgatctg actggcgtgc tctacggaga ggacatcgaa atttctgata cagaaagttt
     2281 ctcgaacgac ccttgcacgt cagtcaaaaa gctgaagggc aacgacgttc gtatcattct
     2341 cggacagttc gatcaaaaca tggcagcgaa ggtgttctgc tgtgcatacg aggaaaacat
     2401 gtacggtagt aaataccagt ggatcattcc cggctggtac gagccatcgt ggtgggaaca
     2461 agtccacacc gaggccaact cttcacgctg tctccgtaag aacctcttgg ctgccatgga
     2521 aggctacatc ggagtggact tcgagccgct gagttcgaag cagatcaaaa ccattagcgg
     2581 aaagactcct cagcaatacg agagggaata caacaacaag agatctggtg tcggcccctc
     2641 aaaattccac ggatacgctt acgacggtat ctgggttatt gcgaagacat tgcaaagggc
     2701 tatggagacg ctgcacgcct ccagcaggca tcagagaatc caagacttca actacacaga
     2761 tcatacgttg ggcagaatca ttctgaacgc catgaacgag acaaacttct tcggtgtcac
     2821 gggccaggtg gtcttcagga acggtgaaag aatgggcaca atcaagttca cgcagttcca
     2881 agactctcgc gaagttaaag tgggagagta caacgcagtg gcggacaccc tggaaatcat
     2941 taacgatact atccgcttcc agggcagtga acccccaaag gacaaaacta tcattctgga
     3001 gcaactccgt aagatctcgt tgcctctgta ctccattttg agcgctctga caatcctcgg
     3061 tatgattatg gcttcagcct tcttgttctt caacatcaag aaccgtaacc agaagctgat
     3121 taaaatgtct tcaccctaca tgaacaactt gatcattctg ggtggtatgc tgtcttacgc
     3181 ctcaatcttc ctcttcggat tggacggtag tttcgtgtcg gagaagacct tcgaaactct
     3241 ctgcacagtc aggacgtgga tcttgacagt tggctacaca acggcattcg gagcgatgtt
     3301 cgctaagacc tggagagtgc acgctatctt caagaacgtc aagatgaaaa agaaaatcat
     3361 taaggaccag aaactgctcg tcatcgttgg cggaatgttg ctgattgacc tgtgcatcct
     3421 catttgttgg caagcagttg atccactgcg ccgtaccgtg gaaaagtact ccatggagcc
     3481 ggaccctgcg ggaagggata tcagcattag acctctcttg gagcactgcg aaaacaccca
     3541 tatgactatc tggctgggta ttgtctacgc ctacaagggt ctgctcatgc tcttcggctg
     3601 tttcttggca tgggagacac gcaacgtgtc tatccccgcg ctgaacgaca gtaaatacat
     3661 tggcatgtcg gtttacaacg tgggaatcat gtgcatcatt ggtgcagcgg tttcattctt
     3721 gactagggat cagccaaacg tgcaattctg tattgtcgct ctggttatca ttttctgcag
     3781 tacaatcacg ctctgtttgg tgttcgtccc gaagttgatc acactgagaa cgaaccctga
     3841 cgctgccacc cagaacagga gattccaatt cactcagaac caaaagaaag aggattccaa
     3901 gacctccact agcgtgacaa gcgtcaacca ggcctctacg tcaaggctcg aaggtttgca
     3961 aagcgagaac cacaggctga gaatgaagat caccgaattg gacaaagatc tggaggaagt
     4021 caccatgcag ctccaagaca cttaataagg cgcgccctcg aggcatgcgg taccaagctt
     4081 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     4141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4321 tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga
     4381 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
     4441 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
     4501 ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt
     4561 ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct
     4621 gtcacagaat gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat
     4681 atcaacgctt atttgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4981 ccctttgacg ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca
     5041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5161 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5221 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5281 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5341 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5401 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5461 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5521 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5581 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5641 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5701 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5761 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5821 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5881 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5941 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6001 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6061 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6121 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7081 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     7141 gtatttcaca ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat
     7201 aaatggatgc cctgcgtaag cggg
//
