LOCUS       sequence-189859-        3017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control gRNA under the regulation of an optimized chicken specific 
            U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3017)
  AUTHORS   Gandhi S, Piacentino ML, Vieceli FM, Bronner ME
  TITLE     Optimization of CRISPR/Cas9 genome editing for loss-of-function in 
            the early chick embryo.
  JOURNAL   Dev Biol. 2017 Dec 1;432(1):86-97. doi: 10.1016/j.ydbio.2017.08.036.
  PUBMED    29150011
REFERENCE   2  (bases 1 to 3017)
  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"; doi: 
            "10.1016/j.ydbio.2017.08"; journalName: "Dev Biol"; date: 
            "2017-12-1- 1"; volume: "432"; issue: "1"; pages: "86-97"
COMMENT     Sequence Label: U6.3>Control.gRNA.f+e
FEATURES             Location/Qualifiers
     source          1..3017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    735..869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(772..791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     826..845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(905..924)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(906..924)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1011..1028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1182..1770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1262..1281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1941..2732,2733..2801))
                     /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     2564..2583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2802..2906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2974..2992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgccgccgc tccttcaggc actgccgacg acagcccagg cggaggtcct gagcgccggc
       61 gctaaatttg cataaagaac tacccaggag ccctcgcgcg cggaaacggg caaaaagggg
      121 cttctaatat ggaaatatta cgccgaatcg cgttacaaat cggctaagcg ggcctaagag
      181 ttaacaagat gtgctattaa gcggagcctt ttggtgggaa gaaatggagt agtcactgtg
      241 ttctaaaaga acttgcagaa tgagccttta aataccgcag tctcgatgct cttagtcgga
      301 tgcactgcta cgatctacac cgtttaagag ctatgctgga aacagcatag caagtttaaa
      361 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgaattc
      421 cagctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata ataataaccg
      481 ggcaggccat gtctgcccgt atttcgcgta aggaaatcca ttatgtacta tttaaaaaac
      541 acaaactttt ggatgttcgg tttattcttt ttcttttact tttttatcat gggagcctac
      601 ttcccgtttt tcccgatttg gctacatgac atcaaccata tcagcaaaag tgatacgggt
      661 attatttttg ccgctatttc tctgttctcg ctattattcc aaccgctgtt tggtctgctt
      721 tctgacaaac tcggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      841 tcatcaatgt atcttatcat gtctggatcg acaaagtcaa agcggccatc agatctgccg
      901 gactccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
      961 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1621 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1921 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1981 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2041 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     2101 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2161 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2221 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2281 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2341 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2401 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2461 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2521 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2581 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2641 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2701 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2761 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2821 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2881 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2941 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcccttacgt
     3001 attaattaag gcgcgcc
//
