LOCUS       sequence-375681-        6517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral delivery of control guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6517)
  AUTHORS   Hosokawa H, Romero-Wolf M, Yui MA, Ungerback J, Quiloan MLG, 
            Matsumoto M, Nakayama KI, Tanaka T, Rothenberg EV
  TITLE     Bcl11b sets pro-T cell fate by site-specific cofactor recruitment 
            and by repressing Id2 and Zbtb16.
  JOURNAL   Nat Immunol. 2018 Dec;19(12):1427-1440. doi: 
            10.1038/s41590-018-0238-4. Epub 2018 Oct 30.
  PUBMED    30374131
REFERENCE   2  (bases 1 to 6517)
  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: 
            "10.1038/s41590-018-0238-4"; journalName: "Nat Immunol"; date: 
            "2018-12"; volume: "19"; issue: "12"; pages: "1427-1440"
COMMENT     Sequence Label: E42_control_NGFR
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(158..175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(329..917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(409..428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1088..1879,1880..1948))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1711..1730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1949..2053)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2121..2139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2239..2618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2619..2822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2773..2793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2823..2998
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3061..3260
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3461..3877
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3807..3829
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3845..3861
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        3893..4141
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3893..3913
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4064..4083
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        4281..4492
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4509..4518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             5533..5958
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6061..6083)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6219..6239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6224..6517,1..36)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6404..6517,1..22)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg
       61 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
      121 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      181 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      241 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      301 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      361 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      421 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      481 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      541 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      601 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      661 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      721 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
      781 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      841 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      901 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
      961 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1021 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1081 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1141 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1201 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1261 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1321 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1381 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1441 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1501 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1561 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1621 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1681 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     1741 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1801 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1861 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1921 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     1981 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2041 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2101 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag
     2161 cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     2221 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     2281 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2341 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2401 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2461 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2521 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2581 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2641 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2701 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2761 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2821 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcttccgata gactgcgtcg
     2881 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggtctcgc
     2941 tgttccttgg gagggtctcc tctgagtgat tgactaccca cgacgggggt ctttcatttg
     3001 ggggctcgtc cgggatttgg agacccctgc ccagggacca ccgacccacc accgggaggt
     3061 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatgt ttgatgttat
     3121 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     3181 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     3241 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     3301 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     3361 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     3421 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     3481 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     3541 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     3601 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     3661 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     3721 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     3781 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     3841 ctccctttat ccagccctca ctccttctct aggcgccgga attgaaggat ccgagggcct
     3901 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     3961 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     4021 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     4081 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     4141 cgggcatttc gcagcctacc ggtttaagag ctatgctgga aacagcatag caagtttaaa
     4201 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctcgag
     4261 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     4321 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4381 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4441 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg
     4501 acgcggtcgc caccatggcc acaaccatgg acgggccgcg cctgctgctg ttgctgcttc
     4561 tgggggtgtc ccttggaggt gccaaggagg catgccccac aggcctgtac acacacagcg
     4621 gtgagtgctg caaagcctgc aacctgggcg agggtgtggc ccagccttgt ggagccaacc
     4681 agaccgtgtg tgagccctgc ctggacagcg tgacgttctc cgacgtggtg agcgcgaccg
     4741 agccgtgcaa gccgtgcacc gagtgcgtgg ggctccagag catgtcggcg ccgtgcgtgg
     4801 aggccgacga cgccgtgtgc cgctgcgcct acggctacta ccaggatgag acgactgggc
     4861 gctgcgaggc gtgccgcgtg tgcgaggcgg gctcgggcct cgtgttctcc tgccaggaca
     4921 agcagaacac cgtgtgcgag gagtgccccg acggcacgta ttccgacgag gccaaccacg
     4981 tggacccgtg cctgccctgc accgtgtgcg aggacaccga gcgccagctc cgcgagtgca
     5041 cacgctgggc cgacgccgag tgcgaggaga tccctggccg ttggattaca cggtccacac
     5101 ccccagaggg ctcggacagc acagccccca gcacccagga gcctgaggca cctccagaac
     5161 aagacctcat agccagcacg gtggcaggtg tggtgaccac agtgatgggc agctcccagc
     5221 ccgtggtgac ccgaggcacc accgacaacc tcatccctgt ctattgctcc atcctggctg
     5281 ctgtggttgt gggccttgtg gcctacatag ccttcaagag gtggaacagt catcgatatc
     5341 ctcgaggtca ccgcggtcta gagtcgacct gcagccaagc ttatcgataa atcgattagt
     5401 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     5461 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     5521 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5581 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     5641 aacagatgga acagggtcga ccctagagaa ccatcagatg tttccagggt gccccaagga
     5701 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5761 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5821 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5881 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     5941 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     6001 ccgacccacc accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa
     6061 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     6181 ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg
     6241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     6421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6481 cgaggccgcc tcggcctctg agctattcca gaagtag
//
