LOCUS       sequence-488809-        8030 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  Fluorescent reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8030)
  AUTHORS   Deglon Lab
  TITLE     Deglon Lab Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVss-Gfa2(B)3-GFP-WPRE-bGH
FEATURES             Location/Qualifiers
     source          1..8030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   23..152
                     /label=AAV2 ITR
     protein_bind    2871..2895
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2963..2972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2969..2971,2972..2974,2975..3685)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2969..2971,2972..2974,2975..3685)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3014..3035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3275..3294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3622..3643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3762..3778)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3789..3813)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3836..3852)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3872..4460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3925..3945)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4343..4354)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4487..4504)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4493..4717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4927..5256)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4941..5076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5241..5261
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5502..6090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5582..5601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6261..7052,7053..7121))
                     /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(7122..7226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7294..7312
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7350..7372)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7472..7491
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7705..7724
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(8014..8030,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8014..8030,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaatacacgg aattaattct agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
       61 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      121 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
      181 cttatctacg tagccatgct ctaggtacca gatctgagct cccacctccc tctctgtgct
      241 gggactcaca gagggagacc tcaggaggca gtctgtccat cacatgtcca aatgcagagc
      301 ataccctggg ctgggcgcag tggcgcacaa ctgtaattcc agcactttgg gaggctgatg
      361 tggaaggatc acttgagccc agaagttcta gaccagcctg ggcaacatgg caagacccta
      421 tctctacaaa aaaagttaaa aaatcagcca cgtgtggtga cacacacctg tagtcccagc
      481 tattcaggag gctgaggtga ggggatcact taaggctggg aggttgaggc tgcagtgagt
      541 cgtggttgcg ccactgcact ccagcctggg caacagtgag accctgtctc aaaagacaaa
      601 aaaaaaaaaa aaaaaaaaaa gaacatatcc tggtgtggag taggggacgc tgctctgaca
      661 gaggctcggg ggcctgagct ggctctgtga gctggggagg aggcagacag ccaggccttg
      721 tctgcaagca gacctggcag cattgggctg gccgcccccc agggcctcct cttcatgccc
      781 agtgaatgac tcaccttggc acagacacaa tgttcggggt gggcacagtg cctgcttccc
      841 gccgcacccc agcccccctc aaatgccttc cgagaagccc attgagcagg gggcttgcat
      901 tgcaccccag cctgacagcc tggcatcttg ggataaaagc agcacagccc cctaggggct
      961 gcccttgctg tgtggcgcca ccggcggtgg agaacaaggc tctattcagc ctgtgcccag
     1021 gaaaggggat caggggatgc ccaggcatgg acagtgggtg gcaggggggg agaggagggc
     1081 tgtctgcttc ccagaagtcc aaggacacaa atgggtgagg ggactgggca gggttctgac
     1141 cctgtgggac cagagtggag ggcgtagatg gacctgaagt ctccagggac aacagggccc
     1201 aggtctcagg ctcctagttg ggcccagtgg ctccagcgtt tccaaaccca tccatcccca
     1261 gaggttcttc ccatctctcc aggctgatgt gtgggaactc gaggaaataa atctccagtg
     1321 ggagacggag gggtggccag ggaaacgggg cgctgcagga ataaagacga gccagcacag
     1381 ccagctcatg tgtaacggct ttgtggagct gtcaaggcct ggtctctggg agagaggcac
     1441 agggaggcca gacaaggaag gggtgacctg gagggacaga tccaggggct aaagtcctga
     1501 taaggcaaga gagtgccggc cccctcttgc cctatcagga cctccactgc cacatagagg
     1561 ccatgattga cccttagaca aagggctggt gtccaatccc agcccccagc cccagaactc
     1621 cagggaatga atgggcagag agcaggaatg tgggacatct gtgttcaagg gaaggactcc
     1681 aggagtctgc tgggaatgag gcctagtagg aaatgaggtg gcccttgagg gtacagaaca
     1741 ggttcattct tcgccaaatt cccagcacct tgcaggcact tacagctgag tgagataatg
     1801 cctgggttat gaaatcaaaa agttggaaag caggtcagag gtcatctggt acagcccttc
     1861 cttccctttt tttttttttt ttttgtgaga caaggtctct ctctgttgcc caggctggag
     1921 tggcgcaaac acagctcact gcagcctcaa cctactgggc tcaagcaatc ctccagcctc
     1981 agcctcccaa agtgctggga ttacaagcat gagccacccc actcagccct ttccttcctt
     2041 tttaattgat gcataataat tgtaagtatt catcatggtc caaccaaccc tttcttgacc
     2101 caccttccta gagagagggt cctcttgctt cagcggtcag ggccccagac ccatggtctg
     2161 gctccaggta ccacctgcct catgcaggag ttggcgtgcc caggaagctc tgcctctggg
     2221 cacagtgacc tcagtggggt gaggggagct ctccccatag ctgggctgcg gcccaacccc
     2281 accccctcag gctatgccag ggggtgttgc caggggcacc cgatcttcct cttcatgccc
     2341 agtgaatgac tcaccttggc acagacacaa tgttcggggt gggcacagtg cctgcttccc
     2401 gccgcacccc agcccccctc aaatgccttc cgagaagccc attgagcagg cccgatcttc
     2461 ctcttcatgc ccagtgaatg actcaccttg gcacagacac aatgttcggg gtgggcacag
     2521 tgcctgcttc ccgccgcacc ccagcccccc tcaaatgcct tccgagaagc ccattgagca
     2581 ggcccgatct tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg
     2641 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa
     2701 gcccattgag caggcccggg catcgccagt ctagcccact ccttcataaa gccctcgcat
     2761 cccaggagcg agcagagcca gagcaggttg gagaggagac gcatcacctc cgctgctcgc
     2821 ggggatcctc tagagtcgac tagtggatcc cccgggctgc aggaattatc acaagtttgt
     2881 acaaaaaagc aggctccacc atgggaacca attcagtcga ctggatccag gcctaagctt
     2941 acgcgtccta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3001 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3061 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3121 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     3181 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3241 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3301 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3361 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3421 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     3481 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3541 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     3601 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3661 tctcggcatg gacgagctgt acaagtccgg actcagatct cgactagcta gtagctagct
     3721 agctagtcga gctcaagctt cgaattcgat atcaagctta tcgataccgt cgacctcgag
     3781 atatctagac ccagctttct tgtacaaagt ggtgataatt cgatatcaag cttatcgata
     3841 ccgtcgacct cgagatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     3901 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3961 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4021 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4081 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4141 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4201 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4261 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4321 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4381 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4441 cctttgggcc gcctccccgc atcgataccg tcgactcgct gatcagcctc gactgtgcct
     4501 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4561 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4621 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4681 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4741 tggggctcga ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     4801 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4861 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca
     4921 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     4981 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5041 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5101 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5161 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5221 ggagcctggg gactttccac accctaactg acacacattc cacagctgca ttaatgaatc
     5281 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5341 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5941 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7261 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7321 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     7381 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     7441 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     7501 caccattcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     7561 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     7621 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     7681 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     7741 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctggc tagcgatgac
     7801 cctgctgatt ggttcgctga ccatttccgg gtgcgggacg gcgttaccag aaactcagaa
     7861 ggttcgtcca accaaaccga ctctgacggc agtttacgag agagatgata gggtctgctt
     7921 cagtaagcca gatgctacac aattaggctt gtacatattg tcgttagaac gcggctacaa
     7981 ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
