LOCUS       sequence-330305-        4896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression of PACmn, a photoactivatable adenylyl 
            cyclase derived from bPAC, membrane-anchored, no/reduced dark 
            activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4896)
  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 4896)
  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-2xLyn-ERex-Venus-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..4896
                     /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"
     regulatory      116..125
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(227..229,230..940)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(269..290)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(530..549)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     877..898
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     3'UTR           2029..2154
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2238..2256)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2239..2256)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2274..2290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2274..2290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2287..2309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2298..2314
                     /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(2322..2328,2329..2346,2347..2352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2367..2388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2505..2522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2676..3264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2756..2775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3435..4226,4227..4295))
                     /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     4058..4077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4296..4400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4468..4486
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4524..4546)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4646..4665
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4859..4881
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4873..4890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4874..4890
                     /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 catgggatgt atcaaatcga aaggtaagga cagcctgtca ggaagtggtt gcattaagag
      181 caagggaaag gattcgttct gctatgaaaa tgaagttgcc agatctgtga gcaagggcga
      241 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      301 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      361 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
      421 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      481 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      541 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      601 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      661 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
      721 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
      781 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
      841 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
      901 cgccgccggg atcactctcg gcatggacga gctgtacaag ctcgagaaga tgaagcggct
      961 ggtgtacatc agcaagatca gcggccacct gagcctggaa gagatccagc ggatcggcaa
     1021 ggtgtccatc aagaacaacc agcgggacaa catcaccggc gtgctgctgt acctgcaggg
     1081 cctgttcttc cagatcctgg aaggcgagaa cgagaaggtg gacaagctgt acaagaaaat
     1141 cctggtggac gaccggcaca ccaacatcct gtgcctgaaa accgagtacg acatcaccga
     1201 ccggatgttc cccaactggg ccatgaaaac catcaacctg aacgagaaca gcgagctgat
     1261 gatccagccc atcaagagcc tgctgcagac catcacccag agccaccggg tgctggaaaa
     1321 gtacatgccc gccagagtga tctacctgat caaccagggc atcaaccccc tgaccgtgga
     1381 gccccagctg gtcgagaaga tcatcttctt cagcgacatc ctggccttca gcaccctgac
     1441 cgagaagctg cccgtgaacg aggtggtcat cctggtcaac cggtacttca gcatctgcac
     1501 ccggatcatc agcgcctacg gcggcgaagt gaccaaatat attggcgatt gcgtgatggc
     1561 cagcttcacc aaagagcagg gcgacgccgc catccggacc agcctggaca tcatctccga
     1621 gctgaagcag ctgcggcacc acgtggaggc caccaacccc ctgcacctgc tgtacaccgg
     1681 catcggcctg agctacggcc acgtgatcga gggcaacatg ggcagcagcc tgaagatgga
     1741 ccacaccctg ctgggggacg ccgtgaacgt ggccgccagg ctggaagccc tgacaagaca
     1801 gctgccctac gccctggcct ttacagccgg cgtgaagaag tgctgccagg cccagtggac
     1861 cttcatcaac ctgggagccc accaggtcaa gggcaagcag gaagccatcg aggtctacac
     1921 cgtgaatgag gcccagaagt actacgacac cctgcagatc acccagctga tccggcagac
     1981 cctggaaaac gacaagtaga ctagttaagc ttgatctggt taccactaaa ccagcctcaa
     2041 gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt
     2101 cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattcta
     2161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aamccccccc cccccccccc tgcaggcatg
     2221 caagctagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag
     2281 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2341 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2401 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2461 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2521 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2581 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3301 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3361 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3421 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3481 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3541 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3601 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3661 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3721 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3781 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3841 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3901 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3961 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4021 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4081 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4141 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4201 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4321 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4381 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4441 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4501 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4561 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4621 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4681 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     4741 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4801 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     4861 cagtcacgac gttgtaaaac gacggccagt gaattg
//
