LOCUS       pAAV-CMVie_synap        5865 bp DNA     circular SYN 05-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5865)
  AUTHORS   Joris VA
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2024 from SnapGene 7.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   1..130
                     /label=5'-ITR
                     /note="/vntifkey=34"
     promoter        211..973
                     /label=CMVie synapsin
                     /label=CMVie\synapsin
                     /note="/vntifkey=29"
     enhancer        271..574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1009..1141
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1178..1187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1184..1894
                     /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"
     misc_feature    1922..1946
                     /label=miR30stemloop sense
     misc_feature    1952..1973
                     /label=miR7 Rn ATP10B
     ncRNA           1976..1990
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    1993..2014
                     /label=miR7 Rn ATP10B
     misc_feature    2016..2039
                     /label=miR30 stem loop asense
     misc_feature    2075..2616
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(2546..2557)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2630..2837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2925..3054
                     /label=3'-ITR
                     /note="/vntifkey=34"
     primer_bind     complement(3073..3089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3231..3686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(3248..3554)
                     /direction=LEFT
                     /label=F1 ori
                     /label=F1\ori
                     /note="/vntifkey=33"
     promoter        3713..3817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3818..4678
                     /codon_start=1
                     /label=Amp-R
                     /note="/vntifkey=4"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4849..5437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5725..5746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5761..5766,5767..5784,5785..5791)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5799..5815
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5823..5839
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctaga tagttattaa tagtaatcaa ttacggggtc
      241 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      301 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      361 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      421 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      481 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      541 gtacatctac gtattagtca tcgctattac catggcacct gcagagggcc ctgcgtatga
      601 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      661 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      721 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      781 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      841 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      901 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      961 gaccatctgc gctgcggccg cactagttgg tcgtgaggca ctgggcaggt aagtatcaag
     1021 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     1081 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     1141 ggtgtccagg cggccgctct agaggatccc caccggtgcc accatggtga gcaagggcga
     1201 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     1261 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1321 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1381 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1441 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1501 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1561 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1621 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1681 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1741 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1801 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1861 ctccaccggc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gctcgagatc
     1921 caagaaggta tattgctgtt gacagtgagc gacctaagac agtgcctata tattagtgaa
     1981 gccacagatg taatatatag gcactgtctt aggatgccta ctgcctcgga cttcaagggc
     2041 tacgatgtcg acttggatcc atcgcgcgtg atccaatcaa cctctggatt acaaaatttg
     2101 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2161 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2221 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2281 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2341 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2401 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2461 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     2521 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2581 cctgctgccg gctctgcggc ctcttccgcg tcttcgagat ctgcctcgac tgtgccttct
     2641 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2701 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2761 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2821 agcaggcatg ctggggactc gagttaaggg cgaattcccg ataaggatct tcctagagca
     2881 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     2941 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3001 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa
     3061 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3121 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3181 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3241 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3301 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3361 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3421 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3481 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3541 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3601 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3661 taacaaaata ttaacgttta taatttcagg tggcatcttt cggggaaatg tgcgcggaac
     3721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3961 tctcaatagt ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagtaatgg taacaacgtt
     4381 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4441 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4501 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4561 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4621 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4681 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4981 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5461 acggttcctg gccttttgct gcggttttgc tcacatgttc tttcctgcgt tatcccctga
     5521 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5581 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5641 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5701 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5761 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5821 cacaggaaac agctatgacc atgattacgc cagatttaat taagg
//
