LOCUS       Exported                6277 bp ds-DNA     circular SYN 06-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-AscI-CAG-NES-StayGold-P2A-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  AUTHORS   Vax
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 6, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6277
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(941..1801)
                     /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(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    2661..2691
                     /label=AscI-OxE insert
                     /label=AscI-OxE\insert
                     /note="/vntifkey=21"
     misc_feature    2692..2935
                     /label=CMV enhancer
                     /label=CMV\enhancer
                     /note="/vntifkey=21"
     promoter        2942..3219
                     /label=chicken beta-actin promoter
     CDS             3643..3678
                     /codon_start=1
                     /label=NES
                     /note="/vntifkey=21"
                     /translation="LQLPPLERLTLS"
     CDS             3685..4335
                     /codon_start=1
                     /label=StayGold
                     /note="/vntifkey=21"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     CDS             4342..4434
                     /codon_start=1
                     /label=P2A
                     /note="/vntifkey=21"
                     /translation="RPVHVGSGATNFSLLKQAGDVEENPGPHVTS"
     CDS             4456..5166
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4497..4516)
                     /label=mCherry-Rseq
     misc_feature    5179..5767
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5650..5661)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5810..5931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(6277,1..940)
                     /label=ColE1
                     /note="ColE1"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa ggcgcgcccc taggggagat ggccagccag cccattgacg
     2701 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2761 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2821 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2881 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2941 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     3001 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     3061 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
     3121 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     3181 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     3241 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     3301 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
     3361 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     3421 ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct gccttcgggg
     3481 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     3541 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     3601 gctgtctcat cattttggca aagaattgga tccgacacca tgcttcaact tcctcctctt
     3661 gaacgtctta ctctttcgag atctatggct agtacaccat ttaaatttca acttaaagga
     3721 accatcaatg gcaaatcgtt taccgttgaa ggcgaaggtg aaggaaattc acatgaaggt
     3781 tctcataaag gaaaatatgt ttgtacaagt ggaaaactac cgatgtcatg ggcagcactt
     3841 ggaacatcct ttggttatgg aatgaaatat tataccaaat atcctagtgg actgaagaac
     3901 tggtttcatg aagtaatgcc tgaaggcttt acctacgatc gtcatattca atataaaggc
     3961 gatgggagta tccatgcaaa acaccaacac tttatgaaaa atgggactta tcacaacatt
     4021 gttgaattta ctggtcagga ttttaaagaa aatagtccag tcttaactgg agatatgaat
     4081 gtctcattac cgaatgaagt tcaacatata cccagagatg atggagtaga atgcccagtg
     4141 accttgcttt atcctttatt atcggataaa tcaaaatgcg ttgaggctca ccaaaataca
     4201 atctgcaagc ctcttcataa tcaaccagca cctgatgtcc catatcactg gattcgtaaa
     4261 caatacacac aaagcaaaga tgataccgag gaacgtgatc atatttgtca atcagagact
     4321 ctcgaagcac acttagaatt ccgaccggtt cacgtgggaa gtggggctac aaacttctca
     4381 ctgctgaaac aggccgggga tgtggaggaa aacccaggac cacacgtgac tagtggtggt
     4441 tctggttctg gtaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     4501 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     4561 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     4621 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     4681 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     4741 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     4801 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     4861 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     4921 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     4981 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     5041 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     5101 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtat
     5161 aagtaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5221 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5281 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5341 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5401 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5461 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5521 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5581 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5641 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5701 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5761 tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     5821 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     5881 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     5941 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     6001 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     6061 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     6121 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     6181 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     6241 caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
