LOCUS       sequence-445242-        6466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of iPASnFR-L from GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  AUTHORS   Tamura R, Chen J, De Jaeger M, Morris JF, Scott DA, Vangheluwe P, 
            Looger LL
  TITLE     Genetically encoded fluorescent sensors for visualizing polyamine 
            levels, uptake, and distribution
  JOURNAL   bioRxiv 2024.08.21.609037
REFERENCE   2  (bases 1 to 6466)
  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: "bioRxiv 
            2024.08.21.609037"
COMMENT     Sequence Label: pAAV-GFAP-iPASnFR-L
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     1923..1944
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2047..2068)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2593..3181
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2646..2666)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3064..3075)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3213..3689
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3412..3431)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3729..3869
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3729..3858
                     /label=AAV2 ITR
     rep_origin      3944..4399
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4031..4050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4241..4262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4416..4435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4535..4557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4595..4613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4681..4785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4786..4854,4855..5646)
                     /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"
     rep_origin      5817..6405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6306..6325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgatcta acatatcctg gtgtggagta ggggacgctg
      181 ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag gcagacagcc
      241 aggccttgtc tgcaagcaga cctggcagca ttgggctggc cgccccccag ggcctcctct
      301 tcatgcccag tgaatgactc accttggcac agacacaatg ttcggggtgg gcacagtgcc
      361 tgcttcccgc cgcaccccag cccccctcaa atgccttccg agaagcccat tgagcagggg
      421 gcttgcattg caccccagcc tgacagcctg gcatcttggg ataaaagcag cacagccccc
      481 taggggctgc ccttgctgtg tggcgccacc ggcggtggag aacaaggctc tattcagcct
      541 gtgcccagga aaggggatca ggggatgccc aggcatggac agtgggtggc agggggggag
      601 aggagggctg tctgcttccc agaagtccaa ggacacaaat gggtgagggg agagctctcc
      661 ccatagctgg gctgcggccc aaccccaccc cctcaggcta tgccaggggg tgttgccagg
      721 ggcacccggg catcgccagt ctagcccact ccttcataaa gccctcgcat cccaggagcg
      781 agcagagcca gagcaggttg gagaggagac gcatcacctc cgctgctcgc ggggtctaga
      841 gtcgacgcca ccatgaacac gttgacggtt tacaattggg gtgattacat cgaccctgca
      901 ttaatccgta agttcgagaa gcaaacgggt ataaaggtta tatatcagac gtttgactcc
      961 aatgaagcaa tgatggcaaa gattgctcaa ggtggagcaa cgttcgacgt agcagttcca
     1021 tctgagtacg cgatctctaa gatgaaggaa gatggattgc tgatcccctt agaccattca
     1081 aaattaccta acctcaaata cattaatccg cgcttcttaa atttaagctt cgaccccgga
     1141 aacagatact ctgtccccta cttctggggt accgtgggga tagtttacaa tcccggcatg
     1201 ttgggtggta agaagatcac ttcttggaac gacttgtggg atccagacct gcgcaatcag
     1261 atcctgcttg ttgacggagc tcgtgcggta atcggaatgg gacttaactc actccactat
     1321 tcactcaacg acaccaataa agagcatctc aagaaggcca aagagaagtt ggacagactc
     1381 gtgccaaatg tgaaagccat agtcggggat gaaatcaaga tgttgctggc gaatgaggaa
     1441 gcagcgatag gagttgtgtg gagtggggac gctgcggaga tcatcgccga aaatgacaag
     1501 ttggattacg tggtgcccaa agagggatcg aacttgtggt tcgacaatat ggtgattcca
     1561 aagaccgcga agaatattga aggagcacat aaattcatta acttcatgct ggacccaaag
     1621 aatgcggcgc agaacgccga gtttgttggc tattcaaccc caaataagaa ggctttggat
     1681 tacatgccta aggaaatcac agagtctaag aggcacacgg tctatatcac cgccgacaag
     1741 cagaagaacg gcatcaaggc gaacttcaag atccacctta acgtggagga cggcagcgtg
     1801 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1861 gacaaccact acctgagcta tcagtccgtg ctgagcaaag accccaacga gaagcgcgat
     1921 cacatggtcc tgctggagtt cgtgaccgcc accgggaact ctctcggcat ggacgagctg
     1981 tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg
     2041 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
     2101 ggcgagggcg atgccaccaa cggcaagcta accctgaagt tcatctgcac caccggcaag
     2161 ctgcccgtgc cctggcccac cctcgtgacc accctgagct acggcgtgca gtgcttcgcc
     2221 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2281 gtccaggagc gcaccatcag cttcaaggac gacggaacct acaagacccg cgccgaggtg
     2341 aagttagagg gcgacaccct ggtgaaccgc atcgagctaa agggcatcga cttcaaggag
     2401 gacggcaaca tcctggggca caagctggag tacaacttta actatcatga caagcggttc
     2461 tatcccgatt tcgactcaat cggcaattta gaagtttaca agaatcttgg aaagcgtatg
     2521 ttggcttact acaatgaact gttcctgcaa tttaagatgc accgctaaga attcgatatc
     2581 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2641 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2701 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2761 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2821 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2881 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2941 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3001 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3061 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3121 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3181 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     3241 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3301 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3361 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3421 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3481 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3541 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3601 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3661 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3721 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3781 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3841 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3901 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3961 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4021 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4081 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4141 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4201 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4261 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4321 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4381 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4441 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4501 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4561 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4621 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4801 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4861 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4921 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4981 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5041 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5101 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5161 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5221 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5281 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5341 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5401 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5461 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5521 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5581 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5641 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5701 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
