pAAV-CAG-monArch-PP-KGC-GFP-Kv2.1-ER2

LOCUS       pAAV-CAG-monArch-PP-KGC-GFP-Kv2.1-ER2             7706 bp DNA     circular SYN 12-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7706)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    25..36
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        40..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        421..696
                     /label=chicken beta-actin promoter
     intron          697..1713
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1721..1740
                     /label=CAG-Fw-Seq
     primer_bind     1750..1795
                     /label=0609-CAG-KpnI-monArch-Fw
     misc_feature    1773
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1774..1779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1780..2538
                     /codon_start=1
                     /label=monArch
                     /translation="MVSIALQAGYDLLGDGRPEILWLGIGTLLILIGTFYFLVRGWGVT
                     NKDAREYYTVTILVSGIAGAAYLLMFFGIGLAEVSVGGEMLVIYYARYAEWLFCTPLLL
                     LDLALLAKVDRVIIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNILTALVLVLWTAYPIIWIIGTEGAGVVGLGIETLLFMVLD
                     VTGKVGFGFILLRSRAILGVTEAPEPSAGAD"
     primer_bind     complement(1817..1859)
                     /label=Archon-Rv-20I
     primer_bind     complement(2517..2565)
                     /label=0609-KGC-PP-monArch-Rv
     primer_bind     2517..2565
                     /label=0609-monArch-PP-KGC-Fw
     CDS             2545..2604
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     complement(2566..2585)
                     /label=KGC-ER2-Rv
     CDS             2605..3318
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     3300..3337
                     /label=Kv2.1-Fw-BsrGI
     CDS             3319..3335
                     /codon_start=1
                     /label=Linker
                     /translation="GGSGG"
     CDS             3337..3531
                     /codon_start=1
                     /label=Kv2.1
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     CDS             3532..3537
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="YK"
     CDS             3538..3558
                     /codon_start=1
                     /label=ER2
                     /translation="FCYENEV"
     primer_bind     complement(3584..3608)
                     /label=0609-CAG-WPREXhoI-ER2-Rv
     intron          3598..3730
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3739..4327
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4360..4471
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4567..4585)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4606..4622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4793..4848
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    5121..5126
                     /label=SDS
     primer_bind     complement(5209..5225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5233..5249
                     /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)."
     promoter        complement(5257..5287)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5302..5323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      5428..5563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5583..5717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5774..6714
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(6715..7575)
                     /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(7576..7680)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagagtcatc ctcatcgtcg acattgatta ttgactagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      481 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      541 cgcgcgccag gcggggcggg gcgggggcga gggcggggcg gggcgaggcg gagaggtgcg
      601 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      661 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      721 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      781 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      841 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      901 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      961 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1021 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1081 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1141 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1201 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1261 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1321 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1381 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1441 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1501 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1561 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1621 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1681 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1741 ctgtctcatc attttggcaa agaattcggt accgccacca tggtttctat cgcactgcag
     1801 gctggttatg acctcctcgg agatggtaga cctgaaatcc tgtggctcgg aattggtaca
     1861 ctgctgatac tgatcggaac cttctacttc ctggttagag ggtggggagt gactaacaag
     1921 gatgcacggg agtactacac agtgaccatc ctggtcagcg gtattgccgg tgcagcttat
     1981 ctgttaatgt tctttggtat cggcctcgca gaggtgtcag ttggaggaga gatgctggtc
     2041 atctactacg caagatacgc agagtggctg ttctgcacac cactgctgct gctggacctg
     2101 gcactcctgg ccaaagttga tagagtgatc atcggcactc tcgttggcgt cgatgctctg
     2161 atgattgtca caggcctgat aggagcactg tcacatactg ccattgccag atactcttgg
     2221 tggctgttct caactatctg tatgatcgtc gttctgtatg tcctggccac ttctctgcgg
     2281 tcagccgcta aggagagggg cccagaggtt gcctcaacct tcaacatcct caccgcactg
     2341 gttctggtcc tgtggactgc ttaccctatc atctggatta tcggcaccga gggtgccgga
     2401 gttgtgggcc tgggaatcga aactctgctg ttcatggtgc tggacgtcac cggcaaggtg
     2461 ggcttcgggt ttatcctgct caggtcaaga gccattctgg gagttacaga agcacctgaa
     2521 ccttcagctg gagctgatcc ccccaagagc aggatcacca gcgagggcga gtacatcccc
     2581 ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac cggggtggtg
     2641 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2701 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatttgcac caccggcaag
     2761 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2821 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2881 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2941 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3001 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3061 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3121 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3181 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3241 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3301 atggacgagc tgtacaaggg tgggagcgga ggtacccaga gccagcctat cctgaacacc
     3361 aaggagatgg cccctcagag caagcctcct gaggagctgg agatgagcag catgcctagc
     3421 cctgtggccc ctctgcctgc caggaccgag ggcgtgatcg acatgaggag catgagcagc
     3481 atcgacagct tcatcagctg cgccaccgac ttccctgagg ccaccaggtt ctacaagttc
     3541 tgctacgaga acgaggtgta atgtacaagt aaagcggccc cggactcgag gccgcaggta
     3601 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     3661 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     3721 ctctccacag gtgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3781 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3841 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3901 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3961 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4021 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4081 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4141 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4201 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4261 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4321 tccccgcctg gaattcgagc tcggtacgat cagcctcgac tgtgccttct agttgccagc
     4381 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4441 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4501 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcccagctt
     4561 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4621 tgtgtgaaat tgttatccgc taattcactc ctcaggtgca ggctgcctat cagaaggtgg
     4681 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcgatct ttttccctct
     4741 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     4801 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     4861 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     4921 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     4981 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     5041 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     5101 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     5161 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     5221 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5281 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5341 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     5401 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5461 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5521 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5581 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5641 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5701 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5761 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5941 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6121 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6361 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6541 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6601 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7261 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7621 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7681 cacatttccc cgaaaagtgc cacctg
//