LOCUS pCAG-QuasAr3-KGC-Citrine-KGCx3-ER2 7634 bp DNA circular SYN 22-APR-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7634)
AUTHORS Kiryl Piatkevich
TITLE Direct Submission
JOURNAL Exported Jun 29, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7634
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 24..35
/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 39..418
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 421..696
/label=chicken beta-actin promoter
intron 697..1708
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1716..1735
/label=CAG-Fw-Seq
regulatory 1769..1778
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 1769..1774
/label=Kozak
CDS 1775..2533
/codon_start=1
/label=QuasAr3
/translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
3'UTR 2285..2287
/label=Mutation
CDS 2540..2599
/codon_start=1
/label=KGC
/translation="KSRITSEGEYIPLDQIDINV"
CDS 2600..3316
/codon_start=1
/product="enhanced variant of YFP (Heikal et al., 2001)"
/label=Citrine
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3317..3373
/codon_start=1
/label=KGC
/translation="SRITSEGEYIPLDQIDINV"
CDS 3374..3433
/codon_start=1
/label=KGC
/translation="KSRITSEGEYIPLDQIDINV"
CDS 3434..3493
/codon_start=1
/label=KGC
/translation="KSRITSEGEYIPLDQIDINV"
CDS 3500..3520
/codon_start=1
/label=ER2
/translation="FCYENEV"
intron 3560..3692
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
misc_feature 3701..4289
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4172..4183)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4322..4433
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
polyA_signal 4725..4779
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5138..5154)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5162..5178
/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(5186..5216)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5231..5252
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5311..5506
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5357..5492
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 5512..5646
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(5885..6473)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter complement(7504..7608)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agagtcatcc tcatcgtcga cattgattat tgactagtta
61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
241 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg
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 tcgctgcgtt gccttcgccc
721 cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
781 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
841 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct
901 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
961 gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
1021 ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc gggggggctg
1081 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
1141 cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct gagcacggcc
1201 cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg
1261 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
1321 gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
1381 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
1441 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
1501 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
1561 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
1621 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
1681 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
1741 tcatcatttt ggcaaagaat tcggtaccgc caccatggta agtatcgctc tgcaggctgg
1801 ttacgaccta ctgggtgacg gcagacctga aaccctgtgg ctgggcatcg gcactctgct
1861 gatgctgatt ggaaccttct actttctggt ccgcggatgg ggagtcaccg ataaggatgc
1921 ccgggaatat tacgctgtga ctatcctggt gtccggaatc gcatccgccg catatctgtc
1981 tatgttcttt ggtatcgggc ttactgaggt gtccgtcggg ggcgaaatgt tggatatcta
2041 ttatgccagg tacgcccagt ggctgtttac caccccactt ctgctgctgc acctggccct
2101 tctcgctaag gtggatcggg tgaccatcgg caccctggtg ggtgtggacg ccctgatgat
2161 cgtcactggc ctcatcggag ccttgagcca cacggccata gccagataca gttggtggtt
2221 gttctctaca atttgcatga tagtggtgct ctatgttctg gctacatccc tgcgatctgc
2281 tgcaagagag cggggccccg aggtggcatc tacctttaac accctgacag ctctggtctt
2341 ggtgctgtgg accgcttacc ctatcctgtg gatcataggc actgagggcg ctggcgtggt
2401 gggcctgggc atcgaaactc tgctgtttat ggtgttggac gtgactgcca aggtcggctt
2461 tggctttatc ctgttgagat cccgggctat tctgggcgac accgaggcac cagaacccag
2521 tgccggtgcc gatacgttga agagtagaat cacaagcgaa ggcgagtaca tccccctgga
2581 tcaaatagac ataaatgtaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
2641 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
2701 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
2761 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt gcttcgcccg
2821 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
2881 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
2941 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
3001 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
3061 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
3121 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
3181 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
3241 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
3301 ggacgagctg tacaagagta gaatcacaag cgaaggcgag tacatccccc tggatcaaat
3361 agacataaat gtaaagagta gaatcacaag cgaaggcgag tacatccccc tggatcaaat
3421 agacataaat gtaaagagta gaatcacaag cgaaggcgag tacatccccc tggatcaaat
3481 agacataaat gtaggtggat tttgttatga gaatgaagtg taatgtacaa gtaaagcggc
3541 cccggactcg aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
3601 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
3661 tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa
3721 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
3781 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
3841 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
3901 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
3961 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
4021 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
4081 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
4141 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
4201 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
4261 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg
4321 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
4381 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
4441 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
4501 tgggaagaca atagcaggcc agcttttgtt ccctttaggg ttaattgcgc gcttggcgct
4561 aatcatggtc atagctgttt ccactcctca ggtgcaggct gcctatcaga aggtggtggc
4621 tggtgtggcc aatgccctgg ctcacaaata ccactgagat cgttttccct ctgccaaaaa
4681 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
4741 tttcattgca atagtgtgtt gaattttttg tgtctctcac tcggaaggac atatgggagg
4801 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
4861 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
4921 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt ttttttatat
4981 tttgtttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
5041 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
5101 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
5161 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
5221 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
5281 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
5341 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
5401 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
5461 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
5521 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
5581 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
5641 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
5701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
6061 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
6301 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
6421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
6481 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
6541 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
6601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
6661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
6721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
6781 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
6841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
6901 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
6961 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
7021 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
7081 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggagcact
7141 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
7201 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
7261 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
7321 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
7381 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
7441 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
7501 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
7561 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
7621 aaaagtgcca cctg
//
