LOCUS       sequence-332576-        2819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone plasmid to generate HH-gRNA-HDV segment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2819)
  AUTHORS   Gandhi S, Li Y, Tang W, Christensen JB, Urrutia HA, Vieceli FM, 
            Piacentino ML, Bronner ME
  TITLE     A single-plasmid approach for genome editing coupled with long-term 
            lineage analysis in chick embryos.
  JOURNAL   Development. 2021 Mar 9. pii: dev.193565. doi: 10.1242/dev.193565.
  PUBMED    33688075
REFERENCE   2  (bases 1 to 2819)
  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. 2021 Mar 9. pii: dev.193565. doi: 10.1242/dev.193565."
COMMENT     Sequence Label: HH-gRNA-HDV-Shuttle-Vector
FEATURES             Location/Qualifiers
     source          1..2819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     268..287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     521..538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     625..644
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        625..643
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    680..814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(704..723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     758..777
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     ncRNA           complement(1135..1202)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     ncRNA           complement(1314..1350)
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     primer_bind     complement(1376..1394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1462..1566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1567..1635,1636..2427)
                     /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(1785..1804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2598..2819,1..367)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
       61 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      541 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      601 caggttaacc tggcttatcg aaattaatac gactcactat agggagtccg gcagatctga
      661 tggccgcttt gactttgtcg atccagacat gataagatac attgatgagt ttggacaaac
      721 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      781 atttgtaacc attataagct gcaataaaca agttccgagt ttgtcagaaa gcagaccaaa
      841 cagcggttgg aataatagcg agaacagaga aatagcggca aaaataatac ccgtatcact
      901 tttgctgata tggttgatgt catgtagcca aatcgggaaa aacgggaagt aggctcccat
      961 gataaaaaag taaaagaaaa agaataaacc gaacatccaa aagtttgtgt tttttaaata
     1021 gtacataatg gatttcctta cgcgaaatac gggcagacat ggcctgcccg gttattatta
     1081 tttttgacac cagaccaact ggtaatggta gcgaccggcg ctcagctgga attcgtccca
     1141 ttcgccatgc cgaagcatgt tgcccagccg gcgccagcga ggaggctggg accatgccgg
     1201 ccaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttatttaaac
     1261 ttgctatgct gtttccagca tagctcttaa actgagaccc tctctcggtc tctgacgagc
     1321 ttactcgttt cgtcctcacg gactcatcag ggcgcgcctt aattaatacg taagggcctc
     1381 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1441 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1501 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1561 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1621 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1681 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1741 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1801 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1861 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1921 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1981 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2041 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2101 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2161 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2221 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2281 ggatctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2341 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2401 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2461 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2521 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaat
//
