LOCUS       sequence-313948-        3064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains tracrRNA-hU6 sequence for double sgRNA cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3064)
  AUTHORS   Y Wang
  TITLE     Yongming Wang lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3064)
  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: 
            "Unpublished"
COMMENT     Sequence Label: hU6-Sa_tracr
FEATURES             Location/Qualifiers
     source          1..3064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     rep_origin      552..1007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(639..658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     849..870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1024..1043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1143..1165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1203..1221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1289..1393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1394..1462,1463..2254)
                     /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(1612..1631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2425..3013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2914..2933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gagaccacgg caggtctcag ttttagtact ctggaaacag aatctactaa
      301 aacaaggcaa aatgccgtgt ttatctcgtc aacttgttgg cgagattttt gcggccgcag
      361 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
      421 gggcgaccaa aggtcgcccg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
      481 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
      541 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      601 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
      661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
      781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
      901 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
      961 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     1021 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     1081 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     1141 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     1201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2101 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3061 tttt
//
