LOCUS       sequence-488811-        6535 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 6535)
  AUTHORS   Deglon Lab
  TITLE     Deglon Lab Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6535)
  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            "Unpublished"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-G1B3-GFP-WPRE-bGH
FEATURES             Location/Qualifiers
     source          1..6535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   23..152
                     /label=AAV2 ITR
     protein_bind    1376..1400
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1468..1477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1474..1476,1477..1479,1480..2190)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1474..1476,1477..1479,1480..2190)
                     /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(1519..1540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1780..1799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2127..2148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2267..2283)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2294..2318)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2341..2357)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2377..2965
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2430..2450)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2848..2859)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2992..3009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2998..3222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3432..3761)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3446..3581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3746..3766
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4007..4595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4087..4106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4766..5557,5558..5626))
                     /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(5627..5731)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5799..5817
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5855..5877)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5977..5996
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6210..6229
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6519..6535,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6519..6535,1)
                     /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 gatctaacat atcctggtgt ggagtagggg
      241 acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg gaggaggcag
      301 acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc ccccagggcc
      361 tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg gggtgggcac
      421 agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa gcccattgag
      481 cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa aagcagcaca
      541 gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca aggctctatt
      601 cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg ggtggcaggg
      661 ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt gaggggagag
      721 ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc agggggtgtt
      781 gccaggggca cccgatcttc ctcttcatgc ccagtgaatg actcaccttg gcacagacac
      841 aatgttcggg gtgggcacag tgcctgcttc ccgccgcacc ccagcccccc tcaaatgcct
      901 tccgagaagc ccattgagca ggcccgatct tcctcttcat gcccagtgaa tgactcacct
      961 tggcacagac acaatgttcg gggtgggcac agtgcctgct tcccgccgca ccccagcccc
     1021 cctcaaatgc cttccgagaa gcccattgag caggcccgat cttcctcttc atgcccagtg
     1081 aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg cttcccgccg
     1141 caccccagcc cccctcaaat gccttccgag aagcccattg agcaggcccg ggcatcgcca
     1201 gtctagccca ctccttcata aagccctcgc atcccaggag cgagcagagc cagagcaggt
     1261 tggagaggag acgcatcacc tccgctgctc gcggggatcc tctagagtcg acggatccgg
     1321 ggaattcgcg gccgcactcg agatctagtg gatcccccgg gctgcaggaa ttatcacaag
     1381 tttgtacaaa aaagcaggct ccaccatggg aaccaattca gtcgactgga tccaggccta
     1441 agcttacgcg tcctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1501 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1561 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1621 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1681 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1741 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1801 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1861 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1921 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1981 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2041 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2101 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2161 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgact agctagtagc
     2221 tagctagcta gtcgagctca agcttcgaat tcgatatcaa gcttatcgat accgtcgacc
     2281 tcgagatatc tagacccagc tttcttgtac aaagtggtga taattcgata tcaagcttat
     2341 cgataccgtc gacctcgaga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2401 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2461 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2521 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2581 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2641 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2701 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2761 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     2821 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2881 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2941 atctcccttt gggccgcctc cccgcatcga taccgtcgac tcgctgatca gcctcgactg
     3001 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3061 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3121 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3181 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3241 ccagctgggg ctcgactaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     3301 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3361 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     3421 gcgcagagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     3481 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     3541 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     3601 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     3661 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     3721 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctgcattaat
     3781 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3961 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4021 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4081 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4141 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4201 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4261 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4381 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4441 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4501 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4621 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5761 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5821 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5881 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5941 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6001 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     6061 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     6121 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6181 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6241 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctggctagcg
     6301 atgaccctgc tgattggttc gctgaccatt tccgggtgcg ggacggcgtt accagaaact
     6361 cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga tgatagggtc
     6421 tgcttcagta agccagatgc tacacaatta ggcttgtaca tattgtcgtt agaacgcggc
     6481 tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
