LOCUS       sequence-241203-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the fluorescent protein mNeonGreen in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  TITLE     AAV expression plasmids for production of mNeonGreen
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5448)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CAG-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1892..2480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1945..1965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2363..2374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2523..2644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2588..2607
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2806..2935
                     /label=AAV2 ITR
     rep_origin      complement(3172..3760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3931..4722,4723..4791))
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4922..5377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5059..5080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5271..5290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc rggsggggsg gggggggggg gggggggggg
      601 sggggsgrgg sggrgrggkg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccgccaccat
     1141 ggtttccaag ggcgaagagg acaatatggc cagcctgcct gccacacacg agctgcacat
     1201 ctttggcagc atcaacggcg tggacttcga catggttgga caaggcaccg gcaatcccaa
     1261 cgacggctac gaggaactga acctgaagtc cacaaagggc gacctgcagt tcagcccttg
     1321 gattctggtg cctcacatcg gctacggctt ccaccagtac ctgccttatc ctgacggcat
     1381 gagccctttt caggccgcca tggtggatgg ctctggctac caggttcacc ggaccatgca
     1441 gtttgaagat ggcgccagcc tgaccgtgaa ctacagatac acatacgagg gcagccacat
     1501 caaaggcgag gcccaagtga aaggcacagg ctttccagcc gatggacccg tgatgaccaa
     1561 tagcctgaca gccgccgact ggtgcagaag caagaaaaca taccccaacg acaagaccat
     1621 catcagcacc ttcaagtggt cctacaccac cggcaacggc aagagataca gaagcaccgc
     1681 cagaaccacc tacaccttcg ccaaacctat ggccgccaac tacctgaaga accagcctat
     1741 gtacgtgttc cgcaagaccg agctgaagca cagcaagaca gaactgaact tcaaagagtg
     1801 gcagaaagcc ttcaccgacg tgatgggcat ggacgagctg tataagtagg ctagcctcga
     1861 ggatatcgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     1921 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1981 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2041 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2101 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2161 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2221 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2281 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2341 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2401 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2461 cctttgggcc gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat
     2521 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2581 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2641 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     2701 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc
     2761 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     2821 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2881 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca
     2941 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4981 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5041 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5101 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5161 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5221 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5281 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     5341 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     5401 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccag
//
