LOCUS       sequence-380415-        3744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor template for CRISPR Genome Editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3744)
  AUTHORS   Sharma S, Mukherjee AK, Roy SS, Bagri S, Lier S, Verma M, Sengupta 
            A, Kumar M, Nesse G, Pandey DP, Chowdhury S
  TITLE     Human telomerase is directly regulated by non-telomeric 
            TRF2-G-quadruplex interaction.
  JOURNAL   Cell Rep. 2021 May 18;35(7):109154. doi: 
            10.1016/j.celrep.2021.109154.
  PUBMED    34010660
REFERENCE   2  (bases 1 to 3744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2021-05-18"; volume: "35(7):109154. doi"; pages: " 
            10.1016/j.celrep.2021.109154"
COMMENT     Sequence Label: CCR5-donor-MCS
FEATURES             Location/Qualifiers
     source          1..3744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1597..1608
                     /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)"
     primer_bind     complement(1762..1779)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1762..1778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1771..1793)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1987..2006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2106..2128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2166..2184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2252..2356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2357..2425,2426..3217)
                     /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(2575..2594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3388..3744,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcag
      661 gcctctgcag tcgacgggcc cgggatccga tacgacgcgg ccgcagatgg tgctttcatg
      721 aattccccca acagagccaa gctctccatc tagtggacag ggaagctagc agcaaaattt
      781 aaatgcattc taggaggctg aggcaggagg atcgcttgag cccaggagtt cgagaccagc
      841 ctgggcaaca tagtgtgatc ttgtatctat aaaaataaac aaaattagct tggtgtggtg
      901 gcgcctgtag tccccagcca cttggagggg tgaggtgaga ggattgcttg agcccgggat
      961 ggtccaggct gcagtgagcc atgatcgtgc cactgcactc cagcctgggc gacagagtga
     1021 gaccctgtct cacaacaaca acaacaacaa caaaaaggct gagctgcacc atgcttgacc
     1081 cagtttctta aaattgttgt caaagcttca ttcactccat ggtgctatag agcacaagat
     1141 tttatttggt gagatggtgc tttcatgaat tcccccaaca gagcacgcgt ctcgagtatc
     1201 ctaggtcaga tctacgcgtg ttaacaagct tctccatcta gtggacaggg aagctagcag
     1261 caaaccttcc cttcactaca aaacttcatt gcttggccaa aaagagagtt aattcaatgt
     1321 agacatctat gtaggcaatt aaaaacctat tgatgtataa aacagtttgc attcatggag
     1381 ggcaactaaa tacattctag gactttataa aagatcactt tttatttatg cacagggtgg
     1441 aacaagatgg attatcaagt gtcaagtcca atctatgaca tcaattatta tacatcggag
     1501 ccctgccaaa aaatcaatgt gaagcaaatc gcagcccgcc tcctgcctcc gctctactca
     1561 ctggtgttca tctttggttt tgtgggcaac atgctggtca tcctcatcct gataaactgc
     1621 aaaaggctga agtttaaaca gatggtgctt tcatgaattc ccccaacaga gccaagctct
     1681 ccatctagtg gacagggaag ctagcagcaa acgccggcgg tcgtcatcta gatgcattcg
     1741 cgaggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1801 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1861 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1921 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1981 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2041 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2101 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2161 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2221 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2281 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2341 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2401 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2461 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2521 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2581 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2641 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2701 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2761 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2821 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2881 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2941 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3001 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3061 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3121 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3181 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3241 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3301 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3721 cacacagccc agcttggagc gaac
//
