LOCUS       sequence-338416-        6938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocyte 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 6938)
  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 6938)
  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: pAAV gfaABC1D 2xLyn-ERex-Venus-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     393..414
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(605..624)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          607..739
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   1544..1673
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1735..2323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1815..1834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2494..3285,3286..3354))
                     /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"
     promoter        complement(3355..3459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3527..3545
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3583..3605)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3705..3724
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3741..4196)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3878..3899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4090..4109
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4271..4411
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4282..4411
                     /label=AAV2 ITR
     polyA_signal    4451..4927
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     4709..4728
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     4994..5010
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4995..5011
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5014..5602
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5120..5131
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5529..5549
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(6681..6938,1..453,454..456))
                     /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(6723..6744)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
       61 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      121 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      181 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      241 ggcgaagcac tgcaggccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
      301 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
      361 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      421 caccaccccg gtgaacagct cctcgccctt gctcacagat ctggcaactt cattttcata
      481 gcagaacgaa tcctttccct tgctcttaat gcaaccactt cctgacaggc tgtccttacc
      541 tttcgatttg atacatccca tggtggcggc cgcgaattca gctgtaattg aactgggagt
      601 ggacacctgt ggagagaaag gcaaagtgga tgtcagtaag accaataggt gcctatcaga
      661 aacgcaagag tcttctctgt ctcgacaagc ccagtttcta ttggtctcct taaacctgtc
      721 ttgtaacctt gatacttacc tgcccagtgc ctcacgacca acttctgcag cttaagttcg
      781 agactgttgt gtcagaagca ctgactgcgt tagcaattta actgtgataa actaccgcaa
      841 taaagcttgc gagcagcgga ggtgatgcgt ctcctctcca acctgctctg gctctgctcg
      901 ctcctgggat gcgagggctt tatgaaggag tgggctagac tggcgatgcc cgggtgcccc
      961 tggcaacacc ccctggcata gcctgagggg gtggggttgg gccgcagccc agctatgggg
     1021 agagctctcc cctcacccat ttgtgtcctt ggacttctgg gaagcagaca gccctcctct
     1081 ccccccctgc cacccactgt ccatgcctgg gcatcccctg atcccctttc ctgggcacag
     1141 gctgaataga gccttgttct ccaccgccgg tggcgccaca cagcaagggc agcccctagg
     1201 gggctgtgct gcttttatcc caagatgcca ggctgtcagg ctggggtgca atgcaagccc
     1261 cctgctcaat gggcttctcg gaaggcattt gaggggggct ggggtgcggc gggaagcagg
     1321 cactgtgccc accccgaaca ttgtgtctgt gccaaggtga gtcattcact gggcatgaag
     1381 aggaggccct ggggggcggc cagcccaatg ctgccaggtc tgcttgcaga caaggcctgg
     1441 ctgtctgcct cctccccagc tcacagagcc agctcaggcc cccgagcctc tgtcagagca
     1501 gcgtccccta ctccacacca ggatatgtta cgcgtgcggc cgcaggaacc cctagtgatg
     1561 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     1621 gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg
     1681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3481 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3541 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3601 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3661 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3721 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     3781 tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa
     3841 atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact
     3901 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3961 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4021 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4081 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
     4141 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
     4201 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4261 atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     4321 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     4381 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac gtggttacct
     4441 acaaaatcag aaggacaggg aagggagcag tggttcacgc ctgtaatccc agcaatttgg
     4501 gaggccaagg tgggtagatc acctgagatt aggagttgga gaccagcctg gccaatatgg
     4561 tgaaaccccg tctctaccaa aaaaacaaaa attagctgag cctggtcatg catgcctgga
     4621 atcccaacaa ctcgggaggc tgaggcagga gaatcgcttg aacccaggag gcggagattg
     4681 cagtgagcca agattgtgcc actgcactcc agcttggttc ccaatagacc ccgcaggccc
     4741 tacaggttgt cttcccaact tgccccttgc tccataccac ccccctccac cccataatat
     4801 tatagaagga cacctagtca gacaaaatga tgcaacttaa ttttattagg acaaggctgg
     4861 tgggcactgg agtggcaact tccagggcca ggagaggcac tggggagggg tcacagggat
     4921 gccacccgta gatcatctct agactcgagg tcgaggccag agaggccatc gaatggcgcg
     4981 ccgcgatcgc gtctcgaggt cgacggtatc gatgcgggga ggcggcccaa agggagatcc
     5041 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
     5101 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     5161 gaatccaggt ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca
     5221 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     5281 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     5341 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     5401 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     5461 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     5521 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     5581 ttttgtaatc cagaggttga ttatcgataa gcttaactag tctacttgtc gttttccagg
     5641 gtctgccgga tcagctgggt gatctgcagg gtgtcgtagt acttctgggc ctcattcacg
     5701 gtgtagacct cgatggcttc ctgcttgccc ttgacctggt gggctcccag gttgatgaag
     5761 gtccactggg cctggcagca cttcttcacg ccggctgtaa aggccagggc gtagggcagc
     5821 tgtcttgtca gggcttccag cctggcggcc acgttcacgg cgtcccccag cagggtgtgg
     5881 tccatcttca ggctgctgcc catgttgccc tcgatcacgt ggccgtagct caggccgatg
     5941 ccggtgtaca gcaggtgcag ggggttggtg gcctccacgt ggtgccgcag ctgcttcagc
     6001 tcggagatga tgtccaggct ggtccggatg gcggcgtcgc cctgctcttt ggtgaagctg
     6061 gccatcacgc aatcgccaat atatttggtc acttcgccgc cgtaggcgct gatgatccgg
     6121 gtgcagatgc tgaagtaccg gttgaccagg atgaccacct cgttcacggg cagcttctcg
     6181 gtcagggtgc tgaaggccag gatgtcgctg aagaagatga tcttctcgac cagctggggc
     6241 tccacggtca gggggttgat gccctggttg atcaggtaga tcactctggc gggcatgtac
     6301 ttttccagca cccggtggct ctgggtgatg gtctgcagca ggctcttgat gggctggatc
     6361 atcagctcgc tgttctcgtt caggttgatg gttttcatgg cccagttggg gaacatccgg
     6421 tcggtgatgt cgtactcggt tttcaggcac aggatgttgg tgtgccggtc gtccaccagg
     6481 attttcttgt acagcttgtc caccttctcg ttctcgcctt ccaggatctg gaagaacagg
     6541 ccctgcaggt acagcagcac gccggtgatg ttgtcccgct ggttgttctt gatggacacc
     6601 ttgccgatcc gctggatctc ttccaggctc aggtggccgc tgatcttgct gatgtacacc
     6661 agccgcttca tcttctcgag cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     6721 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     6781 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     6841 ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg
     6901 aagttggcct tgatgccgtt cttctgcttg tcggcggt
//
