LOCUS       sequence-309786-        7828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HA-TEV-VFT-TMD-CC-PreX-eGFP in pFastBac1 for Sf9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7828)
  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 7828)
  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-10316:GABR1
FEATURES             Location/Qualifiers
     source          1..7828
                     /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             1620..1640
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3906..3929
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3930..3932,3933..3935,3936..4646)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4808..4942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4845..4864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4899..4918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4971..5136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5320..5774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5407..5426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5616..5637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5800..5904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5905..5973,5974..6765)
                     /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(6123..6142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6936..7524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7425..7444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7678..7695
                     /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 catgaaaaca attatagctc tcagctacat attttgtctg gtgttcgcgg
     1621 agaacttgta cttccaggga tcggaacgcc gtgccgtcta catcggagca ctgttcccaa
     1681 tgagtggagg atggccaggt ggacaggctt gtcaaccagc agtcgagatg gcactggaag
     1741 acgttaacag caggagagac atcctcccag attacgagct gaaactcatt caccatgact
     1801 ctaagtgcga tcctggtcag gctaccaagt acctgtacga actgctctac aacgacccta
     1861 tcaagatcat tttgatgccc ggctgttctt cagtcagcac tctggttgct gaggctgcca
     1921 ggatgtggaa cctgatcgtc ctcagttacg gcagttcgtc ccctgcgctg tcgaacaggc
     1981 agagattccc aaccttcttc cgcactcacc cgtcagctac cttgcataac cctactcgtg
     2041 tgaaactgtt cgagaagtgg ggatggaaga aaatcgccac aattcagcaa accactgaag
     2101 tcttcacctc cactttggac gatctggagg aaagggttaa ggaggcaggt atcgaaatta
     2161 ccttcagaca gtccttcttc tccgaccctg ccgttccggt gaagaacctg aaacgccaag
     2221 acgcacgtat cattgtcggt ctcttctacg aaaccgaggc tcgtaaagtg ttctgcgagg
     2281 tctacaagga aagactcttc ggcaagaagt acgtgtggtt cttgatcggc tggtacgccg
     2341 ataactggtt caagatctac gaccctagca ttaactgtac agtggacgaa atgacggagg
     2401 ccgtcgaagg ccacatcaca acggagattg tcatgctgaa ccccgcaaac acccgtagta
     2461 tctcgaacat gacttctcag gagttcgtgg aaaagctgac aaaacgcctc aagcgtcatc
     2521 ccgaggaaac gggaggtttc caagaggcgc cactggctta cgacgccatc tgggcattgg
     2581 cgctggctct caacaagacc tctggcggag gtggccgttc aggtgtcaga ctggaagatt
     2641 tcaactacaa caaccagaca atcacggacc aaatttaccg cgctatgaac agctcttcat
     2701 tcgagggtgt gtccggccac gttgtgttcg atgcgtccgg cagccgtatg gcttggactt
     2761 tgatcgaaca gctgcaagga ggttcctaca agaaaattgg atactacgac agcaccaagg
     2821 acgatttgtc ttggtcaaaa actgataagt ggatcggcgg atcaccaccg gctgaccaga
     2881 ccctcgtcat taaaactttc aggttcctct cccaaaagtt gttcatcagt gtctcggttc
     2941 tcagttcgtt gggcattgtc ctcgctgtcg tttgcttgag tttcaacatc tacaactcgc
     3001 acgttagata cattcagaac tctcaaccca acctgaacaa cctcaccgcc gttggatgct
     3061 cattggcact ggcagcggtg ttcccgctcg gattggacgg ttaccatatc ggtcgcaacc
     3121 agttcccctt cgtgtgtcaa gcccgtctgt ggttgctggg actgggtttc tccctcggct
     3181 acggaagcat gttcactaag atctggtggg ttcacaccgt gttcactaag aaagaggaaa
     3241 agaaagagtg gcgcaaaaca ctggaaccat ggaagctcta cgcgacggtt ggtctcttgg
     3301 tgggcatgga tgtcctgaca ctcgctatct ggcagattgt cgaccctctg cacaggacaa
     3361 tcgagacgtt cgccaaagag gaaccgaagg aagacatcga tgtgtcaatt ttgcctcagc
     3421 tggagcattg ctccagcaga aagatgaaca cttggctggg aatcttctac ggctacaagg
     3481 gactgctctt gctgctcggt atcttcctcg cttacgagac aaaatccgtt agcacggaaa
     3541 agatcaacga tcaccgcgcg gtgggtatgg ctatttacaa cgtggcggtc ttgtgtctga
     3601 tcacagctcc tgtgacgatg attttgtctt cacagcaaga cgctgccttc gccttcgcat
     3661 ctctggctat cgtgttcagt tcgtacatta ctctcgtggt cttgttcgtc cccaagatgc
     3721 gccgtctgat cacacgtggc gagtggcagt cagaagccca agacaccatg aagactggtt
     3781 ccagcaccaa caacaacgag gaagagaaat cccgcttgct ggaaaaggag aaccgtgaac
     3841 tggagaaaat cattgctgag aaggaagaga gggtgagcga attgagacac cagctgcaat
     3901 cccgcctgga ggtgctgttt caggggccta tggtgagcaa gggagaggag ctgttcaccg
     3961 gagtggtgcc tatcctggtg gagctggacg gcgatgtgaa tggccacaag ttttctgtga
     4021 gcggagaggg agagggcgac gcaacctatg gcaagctgac actgaagttc atctgcacca
     4081 caggcaagct gccagtgccc tggcctaccc tggtgaccac actgacatac ggcgtgcagt
     4141 gtttctctcg gtatccagac cacatgaagc agcacgactt cttcaagagc gccatgcccg
     4201 agggctacgt gcaggagagg accatctttt tcaaggacga tggcaactat aagacccgcg
     4261 ccgaggtgaa gtttgagggc gacacactgg tgaaccggat cgagctgaag ggcatcgact
     4321 tcaaggagga tggcaatatc ctgggccaca agctggagta caactataat agccacaacg
     4381 tgtacatcat ggccgataag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4441 atatcgagga cggctccgtg cagctggccg atcactacca gcagaacacc ccaatcggcg
     4501 acggccctgt gctgctgcca gataatcact atctgtccac acagtctgcc ctgagcaagg
     4561 accctaacga gaagcgggat cacatggtgc tgctggagtt cgtgaccgca gcaggaatca
     4621 cactgggaat ggacgagctg tataaataat aaggcgcgcc ctcgaggcat gcggtaccaa
     4681 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
     4741 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4801 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4921 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
     4981 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     5041 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     5101 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     5161 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     5221 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     5281 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     5341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5521 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5581 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
     5641 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5701 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5761 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5821 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5881 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5941 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6001 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6061 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6121 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6181 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6241 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6301 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6361 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6421 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6481 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6541 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6601 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6661 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6721 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6781 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6841 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6901 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6961 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7021 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7081 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7141 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7201 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7261 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7321 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7381 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7441 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7501 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7561 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7621 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7681 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     7741 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     7801 taataaatgg atgccctgcg taagcggg
//
