LOCUS       sequence-129715-        4887 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4887)
  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 4887)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 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..4887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     primer_bind     163..182
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        163..181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        200..238
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     235..259
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      273..282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             279..989
                     /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(424..442)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     692..711
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1014..1602
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1067..1087)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1485..1496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1634..2110
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1833..1852)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2150..2290
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2150..2279
                     /label=AAV2 ITR
     rep_origin      2365..2820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2452..2471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2662..2683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2837..2856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2956..2978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3016..3034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3102..3206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3207..3275,3276..4067)
                     /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(3425..3444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4238..4826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4727..4746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtatcta gataatacga ctcactatag
      181 ggggatccac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa gcagagctcg
      241 tttagtgaac cgtcagatcg cctggaggta ccgccaccat ggtgagcaag ggcgaggagg
      301 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
      361 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
      421 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
      481 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
      541 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
      601 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
      661 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
      721 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
      781 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
      841 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
      901 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
      961 ccggcggcat ggacgagctg tacaagtaag aattcgatat caagcttatc gataatcaac
     1021 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1081 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1141 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1201 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1261 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     1321 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1381 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     1441 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     1501 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     1561 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     1621 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     1681 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     1741 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     1801 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     1861 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     1921 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     1981 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     2041 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     2101 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     2161 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2221 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2281 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2341 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2581 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2701 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2761 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2821 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2881 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2941 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3001 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3061 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3121 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3181 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3241 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3301 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3361 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3421 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3481 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3541 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3601 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3661 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3721 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3781 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3841 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3901 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3961 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4021 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4081 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4141 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4381 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4621 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4801 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4861 ccttttgctg gccttttgct cacatgt
//
