LOCUS       sequence-330308-        5301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression of a photoactivatable adenylyl cyclase 
            derived from bPAC using glycophorin for anchoring to the membrane 
            with reduced dark activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5301)
  AUTHORS   Yang S, Constantin OM, Sachidanandan D, Hofmann H, Kunz TC, 
            Kozjak-Pavlovic V, Oertner TG, Nagel G, Kittel RJ, Gee CE, Gao S
  TITLE     PACmn for improved optogenetic control of intracellular cAMP.
  JOURNAL   BMC Biol. 2021 Oct 18;19(1):227. doi: 10.1186/s12915-021-01151-9.
  PUBMED    34663304
REFERENCE   2  (bases 1 to 5301)
  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: "BMC Biol.";
            date: "2021-10-18"; volume: "19(1):227. doi"; pages: " 
            10.1186/s12915-021-01151-9"
COMMENT     Sequence Label: pGEM-HE-Glyco-Venus-bPAC(S27A)
FEATURES             Location/Qualifiers
     source          1..5301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           51..93
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             join(578..580,581..583,584..1294)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(623..644)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(884..903)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1231..1252
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2381..2410
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     3'UTR           2434..2559
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2643..2661)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2644..2661)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2679..2695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2679..2695)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2692..2714)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2703..2719
                     /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)."
     promoter        complement(join(2727..2733,2734..2751,2752..2757))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2772..2793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2910..2927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3081..3669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3161..3180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3840..4631,4632..4700))
                     /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     4463..4482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4701..4805)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4873..4891
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4929..4951)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5051..5070
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5264..5286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5278..5295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5279..5295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg cttgttcttt
       61 ttgcagaagc tcagaataaa cgctcaactt tggcagatca attccccggg gatccgccac
      121 catgtacggc aagatcatct tcgtgctgct gctgagcgag atcgtgtcca tcagcgccct
      181 gagcaccacc gaggtggcca tgcacacaag caccagcagc tccgtgacca agagctacat
      241 cagcagccag accaacgaca cccacaagag agacacctac gccgccaccc ccagagccca
      301 cgaagtgtcc gaaatcagcg tgcggaccgt gtaccccccc gaggaagaaa caggcgagag
      361 agtgcagctg gcccaccact tcagcgagcc cgagatcacc ctgatcatct ttggcgtgat
      421 ggccggcgtg atcggcacca tcctgctgat ctcttacggc atcagacggc tgatcaagaa
      481 aagccccagc gacgtgaagc ccctgcccag ccctgatacc gatgtgcctc tgagcagcgt
      541 ggaaatcgag aaccccgaga caagcgacca gctcgagatg gtgagcaagg gcgaggagct
      601 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      661 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
      721 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
      781 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      841 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      901 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      961 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1021 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1081 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1141 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     1201 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1261 cgggatcact ctcggcatgg acgagctgta caagggtagc ggtgggctga ggcttaataa
     1321 gatgaagcgg ctggtgtaca tcagcaagat cagcggccac ctgagcctgg aagagatcca
     1381 gcggatcggc aaggtggcca tcaagaacaa ccagcgggac aacatcaccg gcgtgctgct
     1441 gtacctgcag ggcctgttct tccagatcct ggaaggcgag aacgagaagg tggacaagct
     1501 gtacaagaaa atcctggtgg acgaccggca caccaacatc ctgtgcctga aaaccgagta
     1561 cgacatcacc gaccggatgt tccccaactg ggccatgaaa accatcaacc tgaacgagaa
     1621 cagcgagctg atgatccagc ccatcaagag cctgctgcag accatcaccc agagccaccg
     1681 ggtgctggaa aagtacatgc ccgccagagt gatctacctg atcaaccagg gcatcaaccc
     1741 cctgaccgtg gagccccagc tggtcgagaa gatcatcttc ttcagcgaca tcctggcctt
     1801 cagcaccctg accgagaagc tgcccgtgaa cgaggtggtc atcctggtca accggtactt
     1861 cagcatctgc acccggatca tcagcgccta cggcggcgaa gtgaccaagt tcatcggcga
     1921 ctgcgtgatg gccagcttca ccaaagagca gggcgacgcc gccatccgga ccagcctgga
     1981 catcatctcc gagctgaagc agctgcggca ccacgtggag gccaccaacc ccctgcacct
     2041 gctgtacacc ggcatcggcc tgagctacgg ccacgtgatc gagggcaaca tgggcagcag
     2101 cctgaagatg gaccacaccc tgctggggga cgccgtgaac gtggccgcca ggctggaagc
     2161 cctgacaaga cagctgccct acgccctggc ctttacagcc ggcgtgaaga agtgctgcca
     2221 ggcccagtgg accttcatca acctgggagc ccaccaggtc aagggcaagc aggaagccat
     2281 cgaggtctac accgtgaatg aggcccagaa gtactacgac accctgcaga tcacccagct
     2341 gatccgacag accctggaaa acgacaagcc gcggacgtac gagcaaaagc tgatctcgga
     2401 ggaggacctg taagcttgat ctggttacca ctaaaccagc ctcaagaaca cccgaatgga
     2461 gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc
     2521 cattcgtatc tgctcctaat aaaaaaaaag tttcttcaca ttctaaaaaa aaaaaaaaaa
     2581 aaaaaaaaaa aaaaaaammc cccccccccc ccccctgcag gcatgcaagc tagcttgagt
     2641 attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga
     2701 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2761 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2821 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2881 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2941 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3001 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3061 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3121 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3181 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3241 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3301 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3361 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3421 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3481 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3541 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3601 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3661 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3721 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3781 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3841 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3901 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3961 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4021 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4081 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4141 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4201 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4261 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4321 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4381 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4441 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4501 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4561 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4621 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4681 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4741 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4801 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4861 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4921 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4981 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5041 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5101 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     5161 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5221 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5281 aaaacgacgg ccagtgaatt g
//
