LOCUS       sequence-357187-        4679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab11b to knock in 
            SFFV-T2A-Blasticidin resistance-PolyA cassette (for knock out).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4679)
  AUTHORS   Gemperle J, Harrison TS, Flett C, Adamson AD, Caswell PT
  TITLE     On demand expression control of endogenous genes with DExCon, 
            DExogron and LUXon reveals differential dynamics of Rab11 family 
            members.
  JOURNAL   Elife. 2022 Jun 16;11. pii: 76651. doi: 10.7554/eLife.76651.
  PUBMED    35708998
REFERENCE   2  (bases 1 to 4679)
  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: "Elife."; 
            date: "2022-06-16"; volume: "11. pii: 76651. doi"; pages: " 
            10.7554/eLife.76651"
COMMENT     Sequence Label: pJET-64-SFFV-T2A-BlaR-polyA-R11b
FEATURES             Location/Qualifiers
     source          1..4679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             15..68
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             78..476
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(508..525)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    514..738
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(join(1306..1312,1313..1330,1331..1336))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1351..1372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1492..1509)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1663..2251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1743..1762)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2422..3213,3214..3282))
                     /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     3045..3064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3283..3387)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3455..3473
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3779..3797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3847..3865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4188..4595
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 ctagtggcag cggagagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
       61 ccggccctag gctcgagatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
      121 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
      181 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
      241 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
      301 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
      361 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
      421 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaatcta
      481 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
      541 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
      601 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
      661 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
      721 atgcggtggg ctctatgggt cgacggaacc ggtgctggaa gtggtacccg ggacgacgag
      781 tacgactacc tattcaaagg tgcggccggt ggggcacaga cgggcgaagt cgtggcggcg
      841 aggcggcggg cagacgggcc aaggccgcgc tatctttcta gaagatctcc tacaatattc
      901 tcagctgcca tggaaaatcg atgttcttct tttattctct caagattttc aggctgtata
      961 ttaaaactta tattaagaac tatgctaacc acctcatcag gaaccgttgt aggtggcgtg
     1021 ggttttcttg gcaatcgact ctcatgaaaa ctacgagcta aatattcaat atgttcctct
     1081 tgaccaactt tattctgcat tttttttgaa cgaggtttag agcaagcttc aggaaactga
     1141 gacaggaatt ttattaaaaa tttaaatttt gaagaaagtt cagggttaat agcatccatt
     1201 ttttgctttg caagttcctc agcattctta acaaaagacg tctcttttga catgtttaaa
     1261 gtttaaacct cctgtgtgaa attattatcc gctcataatt ccacacatta tacgagccgg
     1321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1381 gcgctcactg ccaattgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2101 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2341 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2401 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2461 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2521 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2581 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2641 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2701 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2761 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2821 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2881 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2941 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3001 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3061 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3121 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3181 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3241 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3421 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccgcccct
     3481 gcagccgaat tatattattt ttgccaaata atttttaaca aaagctctga agtcttcttc
     3541 atttaaattc ttagatgata cttcatctgg aaaattgtcc caattagtag catcacgctg
     3601 tgagtaagtt ctaaaccatt tttttattgt tgtattatct ctaatcttac tactcgatga
     3661 gttttcggta ttatctctat ttttaacttg gagcaggttc cattcattgt ttttttcatc
     3721 atagtgaata aaatcaactg ctttaacact tgtgcctgaa caccatatcc atccggcgta
     3781 atacgactca ctatagggag agcggccgcc agatcttccg gatggctcga gtttttcagc
     3841 aagatataat acgactcact atagggcagt cttccgcccc gccctcttga gctcgcccct
     3901 gataggctgg cgcgtccgtc acttcaaaaa ggtccgcatt ccttccgcct ttctccagga
     3961 caccgagggc gaggagggtg gtaccaagcg gcgcccaccc tcagagcact acttccatct
     4021 ctgattggct tcgctgggtg cccgtcgcta ctccactcgc cgctgatagg ccattcaacc
     4081 gccgctctgc gcggcgccgg ctccgccccc gtcgggtgtt tgtggtgggg ctgcggagtc
     4141 gccgatcccg ccggaagcgc caggacaatg ggatcgggtg ggactaggta acgccatttt
     4201 gcaaggcatg gaaaaatacc aaaccaagaa tagagaagtt cagatcaagg gcgggtacat
     4261 gaaaatagct aacgttgggc caaacaggat atctgcggtg agcagtttcg gccccggccc
     4321 ggggccaaga acagatggtc accgcagttt cggccccggc ccgaggccaa gaacagatgg
     4381 tccccagata tggcccaacc ctcagcagtt tcttaagacc catcagatgt ttccaggctc
     4441 ccccaaggac ctgaaatgac cctgcgcctt atttgaatta accaatcagc ctgcttctcg
     4501 cttctgttcg cgcgcttctg cttcccgagc tctataaaag agctcacaac ccctcactcg
     4561 gcgcgccagt cctccgacag actgagtcgc ccggggggga tctggagctc tcgagaattc
     4621 tcacgcgtct gcaggatatc aagcttgcgg taccgcgggc ccgggatcca tggtgagca
//
