LOCUS       sequence-444510-        4875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4875)
  AUTHORS   Zhang F, Cong L, Lodato S, Kosuri S, Church GM, Arlotta P
  TITLE     Efficient construction of sequence-specific TAL effectors for 
            modulating mammalian transcription.
  JOURNAL   Nat Biotechnol. 2011 Feb;29(2):149-53. doi: 10.1038/nbt.1775
  PUBMED    21248753
REFERENCE   2  (bases 1 to 4875)
  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"; doi: "10.1038/nbt"; 
            journalName: "Nat Biotechnol"; date: "2011-02"; volume: "29"; issue:
            "2"; pages: "149-53"
COMMENT     Sequence Label: pAAV-minCMV-mCherry
FEATURES             Location/Qualifiers
     source          1..4875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     151..170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        151..169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        188..226
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     223..247
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      261..270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             267..977
                     /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(412..430)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     680..699
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1002..1590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1055..1075)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1473..1484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1622..2098
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1821..1840)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2138..2278
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2138..2267
                     /label=AAV2 ITR
     rep_origin      2353..2808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2440..2459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2650..2671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2825..2844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2944..2966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3004..3022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3090..3194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3195..3263,3264..4055)
                     /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     complement(3413..3432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4226..4814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4715..4734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttctaga taatacgact cactataggg ggatccacgt
      181 atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg
      241 tcagatcgcc tggaggtacc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
      301 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      361 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      421 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      481 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      541 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      601 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      661 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
      721 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
      781 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
      841 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
      901 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
      961 acgagctgta caagtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     1021 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1081 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1141 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     1201 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     1261 cctgtcagct cctttccgga actttcgctt tccccctccc tattgccacg gcggaactca
     1321 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1381 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     1441 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     1501 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     1561 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     1621 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     1681 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     1741 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     1801 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     1861 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     1921 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     1981 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     2041 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     2101 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2161 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2221 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2281 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2341 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2401 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2641 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2701 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2821 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2881 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2941 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3421 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3721 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4861 cttttgctca catgt
//
