LOCUS       sequence-240436-        7204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral overexpression of Ccdc134.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7204)
  AUTHORS   Henriksson J, Chen X, Gomes T, Ullah U, Meyer KB, Miragaia R, Duddy 
            G, Pramanik J, Yusa K, Lahesmaa R, Teichmann SA
  TITLE     Genome-wide CRISPR Screens in T Helper Cells Reveal Pervasive 
            Crosstalk between Activation and Differentiation.
  JOURNAL   Cell. 2019 Jan 7. pii: S0092-8674(18)31569-1. doi: 
            10.1016/j.cell.2018.11.044.
  PUBMED    30639098
REFERENCE   2  (bases 1 to 7204)
  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: "Cell. 2019 
            Jan 7. pii: S0092-8674(18)31569-1. doi: 10.1016/j.cell.2018.11.044."
COMMENT     Sequence Label: MSCV-Ccdc134-IRES-BFP
FEATURES             Location/Qualifiers
     source          1..7204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2167..2716
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2311..2328)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2538..2557
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2719..2721,2722..2724,2725..3438)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2764..2785)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     LTR             3599..4113
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4275..4291
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4299..4305,4306..4323,4324..4329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4344..4365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4482..4499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4653..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4733..4752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5412..6203,6204..6272))
                     /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     6035..6054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6273..6377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6445..6463
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6501..6523)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6623..6642
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6836..6858
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6986..7005
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcttaac
     1441 catggacctt cttcagttcc tggcagcctt ctctgtcctc ctgtggcctg gcacggaagt
     1501 cacaggcgcc ctgaagagca ctctggaccc gagcctcaag atttacaaga agatgtttga
     1561 ggtgaagcgg cgagagcagc tgttggcact gaagaacctg gcgcagctga atgacatcca
     1621 ccagcagtac aagatcctgg atgtcatgct taagggactc ttcaaggtgc tggaggactc
     1681 ccgcacagtg ctcattgctg ccgatgtgct tccagacggg ccggtccccc aggatgagaa
     1741 gctgaaggat gctttctcgc acgtggtgga gaacacagcc ttttttggtg acgtcgtgct
     1801 gcgcttccca aagattgtcc accactactt tgaccataat tccaactgga acctcctgat
     1861 ccgctggggc atcagcttct gcaaccagac aggcgtcttt gaccaggggc cccactcacc
     1921 catcctcagc ctgatggccc aggagctggg gatcactgag aaagattctg acttccggaa
     1981 cccgtttaaa acagaccaga cagagtttat tcctagcacc gaccctttcc agaaggcttt
     2041 gagggaagaa gaaaaacgcc gcaagaagga ggagaggcgg aaggagatcc ggaaaggccc
     2101 gaggatttca agatcccagt ctgagttgta gtaggctcga gcgggatcaa ttccgccccc
     2161 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2221 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2281 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2341 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2401 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2461 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2521 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2581 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2641 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2701 gaaaaacacg ataataccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2761 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc
     2821 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2881 cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc
     2941 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3001 caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     3061 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcgtcgactt caaggaggac
     3121 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat ctatatcatg
     3181 gccgtcaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa cgtggaggac
     3241 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3301 ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag
     3361 aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac tctcggcatg
     3421 gacgagctgt acaagtaagc ggccgcgact ctagagtcga cctgcaggca tgcaagcttc
     3481 aggtagccgg ctaacgttaa caaccggtac ctctagaact atagctagca tgcgcaaatt
     3541 taaagcgctg atatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3601 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3661 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3721 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3781 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3841 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3901 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3961 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     4021 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4081 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     4141 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     4201 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     4261 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4321 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4381 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4441 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6421 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6481 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6541 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6601 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6661 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6721 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6781 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6841 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6901 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6961 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     7021 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     7081 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     7141 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     7201 gggg
//
