LOCUS       sequence-236169-        5456 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 5456)
  TITLE     AAV expression plasmids for production of mNeonGreen
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5456)
  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            "Unpublished"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..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(190..778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(270..289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(949..1740,1741..1809))
                     /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     1572..1591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1810..1914)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1940..2395)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2077..2098)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2289..2308
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2467..2596
                     /label=AAV2 ITR
     promoter        2907..3184
                     /label=chicken beta-actin promoter
     primer_bind     3463..3485
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3547..3566
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3599..3608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3860..3869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4358..4946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4411..4431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4829..4840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4989..5110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5000..5019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5054..5073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5272..5401
                     /label=AAV2 ITR
ORIGIN
        1 anngnngnct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
       61 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1921 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1981 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2041 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2101 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2161 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2221 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2281 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2341 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     2401 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2461 cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     2521 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     2581 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
     2641 tgctctagga agagtaccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2701 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2761 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2821 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2881 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     2941 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     3001 agcgatgggg gcgggggggg ggggggggcg cgcgccgggc ggggcggggc ggggcgaggg
     3061 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     3121 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     3181 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     3241 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     3301 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     3361 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc
     3421 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     3481 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     3541 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttggatccgc
     3601 caccatggtt tccaagggcg aagaggacaa tatggccagc ctgcctgcca cacacgagct
     3661 gcacatcttt ggcagcatca acggcgtgga cttcgacatg gttggacaag gcaccggcaa
     3721 tcccaacgac ggctacgagg aactgaacct gaagtccaca aagggcgacc tgcagttcag
     3781 cccttggatt ctggtgcctc acatcggcta cggcttccac cagtacctgc cttatcctga
     3841 cggcatgagc ccttttcagg ccgccatggt ggatggctct ggctaccagg ttcaccggac
     3901 catgcagttt gaagatggcg ccagcctgac cgtgaactac agatacacat acgagggcag
     3961 ccacatcaaa ggcgaggccc aagtgaaagg cacaggcttt ccagccgatg gacccgtgat
     4021 gaccaatagc ctgacagccg ccgactggtg cagaagcaag aaaacatacc ccaacgacaa
     4081 gaccatcatc agcaccttca agtggtccta caccaccggc aacggcaaga gatacagaag
     4141 caccgccaga accacctaca ccttcgccaa acctatggcc gccaactacc tgaagaacca
     4201 gcctatgtac gtgttccgca agaccgagct gaagcacagc aagacagaac tgaacttcaa
     4261 agagtggcag aaagccttca ccgacgtgat gggcatggac gagctgtata agtaggctag
     4321 cctcgaggat atcgaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     4381 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4441 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4501 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4561 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4621 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4681 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4741 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     4801 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4861 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4921 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     4981 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5041 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5101 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     5161 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     5221 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     5281 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5341 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgct
//
