LOCUS       sequence-325764-        4942 bp    DNA     circular SYN 20-JUN-2025
DEFINITION  Direct-expressing SYFP2 AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4942)
  TITLE     Enhancer AAVs from the Allen Institute
REFERENCE   2  (bases 1 to 4942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP11259 - pAAV-eHGT_023h-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(128..144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(128..144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(141..163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    152..168
                     /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(176..182,183..200,201..206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(530..1118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(610..629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1289..2080,2081..2149))
                     /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     1912..1931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2150..2254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2322..2340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2378..2400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2500..2519
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2713..2735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2727..2744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2728..2744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2983..3094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3274..3294
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3387..4097,4098..4100,4101..4103))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3429..3450)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3778..3797
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4037..4058
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4100..4109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgctcgctca ctgaggccgc ccgggcaaag cccgggcggc ctcagtgagc gagcgagcgc
       61 gcagagaggg agtggcccag atccccccgg gaagctttct agatctcgag agcttggcgt
      121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      181 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      241 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2281 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2341 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2401 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2461 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2521 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2581 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2641 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2701 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg tcgacgcatg
     2761 cgggcccgga tctgtccctc tctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     2821 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     2881 ggagtggact agtcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2941 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     3001 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     3061 caaacaacag atggctggca actagaaggc acagtcgctc gagccacgga attgtcagtg
     3121 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3181 gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     3241 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     3301 agaataccag tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt
     3361 cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt ccatgccgag agtgatcccg
     3421 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     3481 agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     3541 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     3601 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
     3661 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     3721 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     3781 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     3841 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     3901 ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     3961 atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     4021 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4081 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcttcga aagatctgga
     4141 tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc
     4201 tctcgtgatg accgggatga aggagccggc ctcctggttt atcacctcac ctctaaatag
     4261 ctcccgggct cggtttctta taaattggta tctcaaacag ggatcacctt cccatcccgg
     4321 agaactggga ggcagagtca ctccgggtca agccagacag ccacaaggag gccgcgtcaa
     4381 cagttctggg gctcactctg taccaggcac cgcacggtgt gttattccat ttcatcctcg
     4441 tcagctctct tatcttcatt ttgtttacag agaaactgag gctgagtgtc actgtgtcac
     4501 tcagctagga agattctaac aggcgtggct ggggccaggg tgcgtctctt cctcaaaggt
     4561 tggagttaaa tggtcctggt tctgactcaa gctgcaaccc ccagaggcag aaggagaggg
     4621 atgcatggag gggaatgtct ttttccacca agcagctgct ccctttcaca agaggccagt
     4681 tatgtaagga aggctgcttc ctctcccctt gctaaatctt ttctagtgca gcataaggcc
     4741 agagctaggg aaagtccaca cgggaataaa aagagcagat tgctaagcac agaagccatt
     4801 tataactaaa acgtctcagc tcagcttagg gacgcgtcct aggtcacgac tccacccctc
     4861 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4921 gccgcccggg caaagcccgg gc
//
