LOCUS       Exported                4247 bp ds-DNA     circular SYN 08-OCT-2021
DEFINITION  Targeting vector to introduce an FKBPV-HA2-P2A-mCherry cassette at 
            the mouse SOX2 locus. Degradation Tag (dTAG) system..
ACCESSION   .
VERSION     .
KEYWORDS    pNQL004-SOX2-FKBPV-HA2-P2A-mCherry targeting construct
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4247)
  AUTHORS   Liu NQ, Maresca M, van den Brand T, Braccioli L, Schijns MMGA, 
            Teunissen H, Bruneau BG, Nora EP, de Wit E
  TITLE     WAPL maintains a cohesin loading cycle to preserve 
            cell-type-specific distal gene regulation.
  JOURNAL   Nat Genet. 2021 Jan;53(1):100-109. doi: 10.1038/s41588-020-00744-4. 
            Epub 2020 Dec 14.
  PUBMED    33318687
REFERENCE   2  (bases 1 to 4247)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4247
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     23..40
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    157..178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        193..223
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    231..247
                     /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)."
     primer_bind     236..258
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     255..271
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     255..271
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             517..837
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             844..870
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             874..900
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             910..966
                     /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"
     CDS             967..1674
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1109..1127)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1902..1919)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1902..1918)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1911..1933)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2127..2146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2246..2268
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2306..2324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2392..2496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2497..3357
                     /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(2715..2734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3528..4116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4017..4036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
       61 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      121 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      181 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      241 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
      301 ggtcgacgcc cccccgtggt tacctcttcc tcccactcca gggcgccctg ccaggccggg
      361 gacctccggg acatgatcag catgtacctc cccggcgccg aggtgccgga gcccgctgcg
      421 cccagtagac tgcacatggc ccagcactac cagagcggcc cggtgcccgg cacggccatt
      481 aacggcacac tgcccctgtc gcacatggga tccggaggag tgcaggtgga aaccatctcc
      541 ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg
      601 atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc ctttaagttt
      661 atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg
      721 ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca
      781 ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaggc
      841 ggctacccct acgacgtgcc cgactacgcc ggctatccgt atgatgtccc ggactatgca
      901 ggctccggag caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct
      961 ggaccggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1021 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1081 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1141 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1201 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1261 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1321 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1381 gtaatgcaga agaaaaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1441 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1501 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1561 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1621 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagggctg
     1681 gactgcgaac tggagaaggg gagagatttt caaagagata caagggaatt gggaggggtg
     1741 caaaaagagg agagtaggaa aaatctgata atgctcaaaa ggaaaaaaaa tctccgcagc
     1801 gaaacgacag ctgcggaaaa aaaccaccaa tcccatccaa attaacgcaa aaaccgtgat
     1861 gccgactaga aaacggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa
     1921 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1981 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2041 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2101 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2161 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2221 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2281 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2341 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2461 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2521 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2581 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2641 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2701 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2761 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2821 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2881 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2941 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3001 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3061 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3121 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3181 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3241 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3301 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3361 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3421 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3481 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3661 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4141 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4201 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcg
//
