pGL3-FANCF-Nicking/PegRNA-PGK-puroR-mRFP1

LOCUS       pGL3-FANCF-Nicking/PegRNA-PGK-puroR-mRFP1               5679 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 5679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..55
                     /label=OL2640
     primer_bind     complement(21..44)
                     /label=OL459
     primer_bind     22..61
                     /label=OL2721
     primer_bind     24..40
                     /label=OL1149
     primer_bind     24..40
                     /label=OL2855
     primer_bind     36..60
                     /label=OL535
     misc_feature    41..115
                     /label=hCtRNA
     primer_bind     56..79
                     /label=OL464
     primer_bind     complement(96..115)
                     /label=OL536
     primer_bind     complement(96..115)
                     /label=OL538
     primer_bind     complement(98..115)
                     /label=OL465
     primer_bind     complement(101..116)
                     /label=OL2651
     primer_bind     complement(101..115)
                     /label=OL2653
     misc_feature    116..135
                     /label=FANCF-nickingRNA
     primer_bind     133..159
                     /label=OL537
     primer_bind     135..176
                     /label=OL462
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     136..164
                     /label=OL2726
     primer_bind     136..151
                     /label=OL2652
     primer_bind     complement(194..230)
                     /label=OL463
     misc_feature    212..286
                     /label=hCtRNA
     primer_bind     212..232
                     /label=OL2721
     primer_bind     212..231
                     /label=OL535
     primer_bind     212..226
                     /label=OL2640
     primer_bind     227..250
                     /label=OL464
     primer_bind     complement(267..286)
                     /label=OL536
     primer_bind     complement(267..286)
                     /label=OL538
     primer_bind     complement(269..286)
                     /label=OL465
     primer_bind     complement(272..287)
                     /label=OL2651
     primer_bind     complement(272..286)
                     /label=OL2653
     misc_feature    287..306
                     /label=FANCF-spacer
     misc_RNA        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     307..347
                     /label=OL462
     primer_bind     307..335
                     /label=OL2726
     primer_bind     307..330
                     /label=OL537
     primer_bind     307..322
                     /label=OL2652
     primer_bind     complement(365..385)
                     /label=OL463
     misc_feature    383..399
                     /label=RT template
     primer_bind     complement(392..421)
                     /label=OL550
     misc_feature    395
                     /label=G to T
     misc_feature    400..412
                     /label=PBS
     primer_bind     complement(406..424)
                     /label=OL1150
     primer_bind     complement(413..436)
                     /label=OL2655
     primer_bind     complement(413..436)
                     /label=OL2727
     primer_bind     418..437
                     /label=OL458
     primer_bind     complement(419..435)
                     /label=OL2856
     misc_feature    456..573
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        622..1132
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1051..1069
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1154..1750
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1154..1173)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1650..1670
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1721..1775
                     /label=OL901
     primer_bind     complement(1736..1758)
                     /label=OL384
     primer_bind     1743..1774
                     /label=OL379
     CDS             1751..1807
                     /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"
     primer_bind     1751..1807
                     /label=OL866
     primer_bind     complement(1751..1807)
                     /label=OL867
     primer_bind     1788..1831
                     /label=OL902
     primer_bind     complement(1788..1815)
                     /label=OL966
     RBS             1790..1797
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1808..2485
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2463..2510)
                     /label=OL903
     primer_bind     2484..2502
                     /label=OL965
     polyA_signal    2632..2753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2643..2662)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2697..2716
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3001..3018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3172..3760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3931..4791)
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4923..5378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5010..5029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5220..5241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5509..5557
                     /note="synthetic polyadenylation signal"
     misc_feature    5571..5662
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5611..5630
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
       61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctggggt
      121 cccaggtgct gacgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
      241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctggaa tcccttctgc
      301 agcaccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gcggaaaagc gatcaaggtg ctgcagaagg gatttttttc
      421 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
      481 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      541 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
      601 ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt
      661 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
      721 gactcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
      781 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
      841 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
      901 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
      961 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     1021 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
     1081 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc
     1141 accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1201 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     1261 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1321 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     1381 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     1441 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     1501 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     1561 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     1621 gcccgccttc ctggaaacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     1681 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     1741 gcccggtgcc gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc
     1801 tggacctatg gcgagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat
     1861 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
     1921 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
     1981 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
     2041 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
     2101 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
     2161 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
     2221 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
     2281 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
     2341 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
     2401 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
     2461 aggtcgtcac tccaccggtg cttaacgccc gccccacgac ccgcagcgcc cgaccgaaag
     2521 gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga
     2581 tcttagccac tttctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     2641 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2701 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2761 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2821 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     2881 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2941 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gggacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5281 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5341 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5461 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     5521 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     5581 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     5641 gcaggtgcca gaacatttct ctatcgatag gtaccgatt
//