LOCUS       sequence-191866-        3513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chick-specific U6 sgRNA expression mini-vector, harbouring chick 
            U6_3 pol III promoter, with tracrRNA scaffold containing stem loops 
            allowing binding of bacteriophage MS2 coat protein MCP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3513)
  AUTHORS   Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, 
            Sauka-Spengler T
  TITLE     Genome and epigenome engineering CRISPR toolkit for in vivo 
            modulation of cis-regulatory interactions and gene expression in the
            chicken embryo.
  JOURNAL   Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 
            10.1242/dev.160333.
  PUBMED    29386245
REFERENCE   2  (bases 1 to 3513)
  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: 
            "Development."; date: "2018-02-23"; volume: "145(4). pii: 
            dev.160333. doi"; pages: " 10.1242/dev.160333"
COMMENT     Sequence Label: pcU6_3 MS2 sgRNA
FEATURES             Location/Qualifiers
     source          1..3513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(927..954)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1046..1132)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1356..1361,1362..1378,1379..1384)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1432..2622
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1784..1803)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2796..2815)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2797..2815)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2820..2836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2820..2836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        3283..3301
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3353..3371
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      841 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      901 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      961 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     1021 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     1081 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     1141 cagttcccta ctctcgcgtt ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1201 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1261 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1321 ataacgcagg aaagaacatg aattaattct catgtttgac agcttatcat cgattagctt
     1381 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
     1441 aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg
     1501 gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt
     1561 cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc
     1621 ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc
     1681 actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggattctcta cgccggacgc
     1741 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     1801 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     1861 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttaca tgcaccattc
     1921 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1981 tcgcataagg gagagcgccg acccatgccc ttgagagcct tcaacccagt cagctccttc
     2041 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     2101 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     2161 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     2221 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     2281 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     2341 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     2401 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     2461 accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     2521 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     2581 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     2641 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     2701 atgcgcaaac caacccttga tcggggaaga acagtatgtc gagctatttt ttgacttact
     2761 ggggatcaag cctgattggg agaaaataaa atatccccta tagtgagtcg tattacatgg
     2821 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2881 aagataaaaa tatatcatca tgcctcctct agtatcatca tgcctcctct agctagaggt
     2941 ctcaggaccg cggccgctcc ttcaggcact gccgacgaca gcccaggcgg aggtcctgag
     3001 cgccggcgcc aaatttgcat aaagagctac ccagaagccc tcgcgcgcgg aaacgggcaa
     3061 aaaagggctt ctaatatgga aatattacgc cgaatcgcgt cacaaattgg ctaagcgggc
     3121 ctaagagtta acaagatgtg ctattaagcg gagccttttg gtgggaagaa acggaacagt
     3181 cactgtgttc taaaagaact tgcaggatga gcctttaaat accgcagtct cgacgctctt
     3241 agtcggagac gttgatttcg tctctgtttt agagctaggc caacatgagg atcacccatg
     3301 tctgcagggc ctagcaagtt aaaataaggc tagtccgtta tcaacttggc caacatgagg
     3361 atcacccatg tctgcagggc caagtggcac cgagtcggtg ctttttttga attctgatgc
     3421 ggtattttct cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     3481 cactgagcgt cagaccccgt agaaaagatc aaa
//
