pGL3-GATA2-Nicking/EpegRNA-PGK-puroR-mRFP1

LOCUS       pGL3-GATA2-Nicking/EpegRNA-PGK-puroR-mRFP1             5722 bp DNA     circular SYN 07-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..44)
                     /label=OL5018
     misc_feature    41..115
                     /label=hCtRNA
     misc_feature    116..135
                     /label=GATA2 nickingRNA
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    212..286
                     /label=hCtRNA
     misc_feature    287..306
                     /label=GATA2 spacer
     misc_RNA        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    383..394
                     /label=RTT
     misc_feature    395..410
                     /label=PBS
     misc_feature    411..418
                     /label=Linker
     misc_feature    419..455
                     /label=motif
     primer_bind     419..441
                     /label=OL5073
     misc_feature    499..616
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        665..1175
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1197..1793
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             1794..1850
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     RBS             1833..1840
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1851..2528
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     polyA_signal    2675..2796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3215..3803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3974..4834)
                     /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(4835..4939)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4966..5421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    5552..5600
                     /note="synthetic polyadenylation signal"
     misc_feature    5614..5705
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
       61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctacatc
      121 ttccggttct gagtcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
      241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctagaa ggaagggatc
      301 cagactgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gctcttccgg ttctgagtct ggatcccttc aacaaacgcg
      421 cggttctatc tagttacgcg ttaaaccaac tagaattttt ttctcgacct cgagacaaat
      481 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      541 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      601 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggccgcggct
      661 cgagggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct
      721 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggactcgc acattcttca
      781 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc
      841 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac
      901 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg
      961 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga
     1021 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt
     1081 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct
     1141 ccctcgttga ccgaatcacc gacctctctc cccaggggga tccaccggag cttaccatga
     1201 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     1261 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     1321 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     1381 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     1441 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     1501 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     1561 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     1621 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaaa
     1681 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     1741 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccgctacta
     1801 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct atggcgagta
     1861 gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt tccgttaacg
     1921 gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt acccagaccg
     1981 ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc ctgtccccgc
     2041 agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg gactacctga
     2101 aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa gacggtggtg
     2161 ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac aaagttaaac
     2221 tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc atgggttggg
     2281 aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa atcaaaatgc
     2341 gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc tacatggcta
     2401 aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac atcacctccc
     2461 acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt cactccaccg
     2521 gtgcttaacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     2581 catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttctag
     2641 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2701 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2761 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2821 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2881 tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag
     2941 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     3001 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     3061 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3121 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3841 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3901 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3961 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4021 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4081 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
     4141 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4201 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4261 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4321 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4381 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4441 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4501 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4561 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4621 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4681 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4741 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4801 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4861 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4921 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     4981 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5041 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5101 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5161 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5221 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5281 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5341 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5401 aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc
     5461 gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata
     5521 ttaaggtacg ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct
     5581 gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac
     5641 gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt
     5701 tctctatcga taggtaccga tt
//