LOCUS       sequence-356073-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFastBac1 vector with GP64 promoter replacing polH promoter 
            expressing A2aAR+BRIL GPCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Liu W, Chun E, Thompson AA, Chubukov P, Xu F, Katritch V, Han GW, 
            Roth CB, Heitman LH, IJzerman AP, Cherezov V, Stevens RC
  TITLE     Structural basis for allosteric regulation of GPCRs by sodium ions.
  JOURNAL   Science. 2012 Jul 13;337(6091):232-6. doi: 10.1126/science.1219218.
  PUBMED    22798613
REFERENCE   2  (bases 1 to 6056)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2012-07-13- 13"; volume: "337"; 
            issue: "6091"; pages: "232-6"
COMMENT     Sequence Label: IMPT-2070
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1346..1375
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    1508..1642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1545..1564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1599..1618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1671..1836
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2020..2474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2107..2126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2316..2337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2500..2604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2605..2673,2674..3465)
                     /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(2823..2842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3636..4224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4125..4144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4378..4395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4529..4753
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4820..5353)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5523..5528,5529..5545,5546..5551))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        join(5965..6056,1..8)
                     /gene="Autographa californica gp64"
                     /label=gp64 promoter
                     /note="promoter for the baculovirus gp64 glycoprotein gene"
ORIGIN
        1 caagcacggg cttctcctca aaaggcctgg atccatgaag acgatcatcg ccctgagcta
       61 catcttctgc ctggtgttcg ccgactacaa ggacgatgat gacggcgcgc cacccatcat
      121 gggctcctcg gtgtacatca cggtggagct ggccattgct gtgctggcca tcctgggcaa
      181 tgtgctggtg tgctgggccg tgtggctcaa cagcaacctg cagaacgtca ccaactactt
      241 tgtggtgtca ctggcggcgg ccgacatcgc agtgggtgtg ctcgccatcc cctttgccat
      301 caccatcagc accgggttct gcgctgcctg ccacggctgc ctcttcattg cctgcttcgt
      361 cctggtcctc acgcagagct ccatcttcag tctcctggcc atcgccattg accgctacat
      421 tgccatccgc atcccgctcc ggtacaatgg cttggtgacc ggcacgaggg ctaagggcat
      481 cattgccatc tgctgggtgc tgtcgtttgc catcggcctg actcccatgc taggttggaa
      541 caactgcggt cagccaaagg agggcaagaa ccactcccag ggctgcgggg agggccaagt
      601 ggcctgtctc tttgaggatg tggtccccat gaactacatg gtgtacttca acttctttgc
      661 ctgtgtgctg gtgcccctgc tgctcatgct gggtgtctat ttgcggatct tcctggcggc
      721 gcgacgacag ctggctgatc tggaagacaa ttgggaaact ctgaacgaca atctcaaggt
      781 gatcgagaag gctgacaatg ctgcacaagt caaagacgct ctgaccaaga tgagggcagc
      841 agccctggac gctcagaagg ccactccacc taagctcgag gacaagagcc cagatagccc
      901 tgaaatgaaa gactttcggc atggattcga cattctggtg ggacagattg atgatgcact
      961 caagctggcc aatgaaggga aagtcaagga agcacaagca gccgctgagc agctgaagac
     1021 cacccggaat gcatacattc agaagtacct ggagcgcgct cggtccacac tgcagaagga
     1081 ggtccatgct gccaagtcac tggccatcat tgtggggctc tttgccctct gctggctgcc
     1141 cctacacatc atcaactgct tcactttctt ctgccccgac tgcagccacg cccctctctg
     1201 gctcatgtac ctggccatcg tcctctccca caccaattcg gttgtgaatc ccttcatcta
     1261 cgcctaccgt atccgcgagt tccgccagac cttccgcaag atcattcgca gccacgtcct
     1321 gaggcagcaa gaacctttca aggcacacca ccatcaccat caccatcacc atcactgaaa
     1381 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
     1441 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1501 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1621 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
     1681 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     1741 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     1801 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     1861 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     1921 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     1981 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     2041 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2101 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2161 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2221 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2281 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
     2341 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2401 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2461 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2521 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2581 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2641 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2701 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2761 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2821 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2881 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2941 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3001 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3061 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3121 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3181 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3241 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3301 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3361 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3421 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4321 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4381 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4441 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     4501 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
     4561 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
     4621 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
     4681 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
     4741 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
     4801 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     4861 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     4921 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     4981 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     5041 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     5101 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     5161 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     5221 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     5281 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     5341 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     5401 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     5461 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     5521 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
     5581 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     5641 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     5701 tctacggcaa ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc
     5761 cgtcgcggcg cttgtcggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     5821 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     5881 gtaatcccgt ttttataaca gccagataaa aataatctta tcaattaaga taaaaagata
     5941 agattattaa tctaacaacg tgccttgtgt cacgtaggcc agataacggt cgggtatata
     6001 agacgcctca acgctactag taaatcagtc acaccaaggc ttcaataagg aacaca
//
