LOCUS       sequence-388974-        7858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse CD4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7858)
  AUTHORS   Akama-Garren EH, Yin X, Prestwood TR, Ma M, Utz PJ, Carroll MC
  TITLE     T cell help shapes B cell tolerance.
  JOURNAL   Sci Immunol. 2024 Feb 16;9(92):eadj7029. doi: 
            10.1126/sciimmunol.adj7029. Epub 2024 Feb 16.
  PUBMED    38363829
REFERENCE   2  (bases 1 to 7858)
  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: "Sci 
            Immunol."; date: "2024-02-16"; volume: "9(92):eadj7029. doi"; pages:
            " 10.1126/sciimmunol.adj7029. Epub 2024 Feb 16"
COMMENT     Sequence Label: pMIAII-Cd4
FEATURES             Location/Qualifiers
     source          1..7858
                     /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    2821..3370
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2965..2982)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3192..3211
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3373..3375,3376..3378,3379..4092)
                     /codon_start=1
                     /product="photostable yellow fluorescent protein with a 
                     large Stokes shift, excitable with violet light (Ai et al.,
                     2008)"
                     /label=mAmetrine1.1
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVRGEGEGDATNGKLTL
                     KFICTSGELPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GSYRTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNMNAWDAYITADKQKNGIK
                     ANFKIEHNVKDGGVQLADAYQQNTPIGDGSVLLPDNHYLSFQSKLFKDPNEQRDHMVLL
                     EFVTAAGITPGMDELYK"
     primer_bind     complement(3418..3439)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     LTR             4253..4767
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4929..4945
                     /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(4953..4959,4960..4977,4978..4983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4998..5019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5136..5153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5307..5895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5387..5406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6066..6857,6858..6926))
                     /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     6689..6708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6927..7031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7099..7117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7155..7177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7277..7296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7490..7512
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7640..7659
                     /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 tcaccatgtg ccgagccatc tctcttaggc
     1441 gcttgctgct gctgctgctg cagctgtcac aactcctagc tgtcactcaa gggaagacgc
     1501 tggtgctggg gaaggaaggg gaatcagcag aactgccctg cgagagttcc cagaagaaga
     1561 tcacagtctt cacctggaag ttctctgacc agaggaagat tctggggcag catggcaaag
     1621 gtgtattaat tagaggaggt tcgccttcgc agtttgatcg ttttgattcc aaaaaagggg
     1681 catgggagaa aggatcgttt cctctcatca tcaataaact taagatggaa gactctcaga
     1741 cttatatctg tgagctggag aacaggaaag aggaggtgga gttgtgggtg ttcaaagtga
     1801 ccttcagtcc gggtaccagc ctgttgcaag ggcagagcct gaccctgacc ttggatagca
     1861 actctaaggt ctctaacccc ttgacagagt gcaaacacaa aaagggtaaa gttgtcagtg
     1921 gttccaaagt tctctccatg tccaacctaa gggttcagga cagcgacttc tggaactgca
     1981 ccgtgaccct ggaccagaaa aagaactggt tcggcatgac actctcagtg ctgggttttc
     2041 agagcacagc tatcacggcc tataagagtg agggagagtc agcggagttc tccttcccac
     2101 tcaactttgc agaggaaaac gggtggggag agctgatgtg gaaggcagag aaggattctt
     2161 tcttccagcc ctggatctcc ttctccataa agaacaaaga ggtgtccgta caaaagtcca
     2221 ccaaagacct caagctccag ctgaaggaaa cgctcccact caccctcaag ataccccagg
     2281 tctcgcttca gtttgctggt tctggcaacc tgactctgac tctggacaaa gggacactgc
     2341 atcaggaagt gaacctggtg gtgatgaaag tggctcagct caacaatact ttgacctgtg
     2401 aggtgatggg acctacctct cccaagatga gactgaccct gaagcaggag aaccaggagg
     2461 ccagggtctc tgaggagcag aaagtagttc aagtggtggc ccctgagaca gggctgtggc
     2521 agtgtctact gagtgaaggt gataaggtca agatggactc caggatccag gttttatcca
     2581 gaggggtgaa ccagacagtg ttcctggctt gcgtgctggg tggctccttc ggctttctgg
     2641 gtttccttgg gctctgcatc ctctgctgtg tcaggtgccg gcaccaacag cgccaggcag
     2701 cacgaatgtc tcagatcaag aggctcctca gtgagaagaa gacctgccag tgcccccacc
     2761 ggatgcagaa gagccataat ctcatctgac tcgagcggga tcaattccgc ccccccccta
     2821 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2881 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2941 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     3001 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     3061 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     3121 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     3181 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     3241 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     3301 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     3361 cacgataata ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3421 gtcgagctgg acggcgacgt aaacggccac aggttcagcg tgcgcggcga gggcgagggc
     3481 gatgccacca acggcaagct gaccctgaag ttcatctgca cctccggcga gctgcccgtg
     3541 ccctggccca ccctcgtgac caccctgtct tacggcgtgc agtgcttcgc ccgctacccc
     3601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3661 cgcaccatct ccttcaagga cgacggcagc tacaggaccc gcgccgaggt gaagttcgag
     3721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3781 atcctggggc acaagctgga gtacaacatg aacgcgtggg acgcgtatat cacggccgac
     3841 aagcagaaga acggcatcaa agcgaacttc aagatcgagc acaacgtcaa ggacggcggc
     3901 gtgcagctcg ccgacgcgta ccagcagaac acccccatcg gcgacggctc cgtgctgctg
     3961 cctgacaacc actacctgag cttccagagc aagctgttca aagaccccaa cgagcagcgc
     4021 gatcacatgg tcctgctgga gttcgttacc gccgccggga taactcccgg catggacgag
     4081 ctgtacaagt aagcggccgc gactctagag tcgacctgca ggcatgcaag cttcaggtag
     4141 ccggctaacg ttaacaaccg gtacctctag aactatagct agcatgcgca aatttaaagc
     4201 gctgatatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     4261 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     4321 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     4381 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4441 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     4501 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     4561 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     4621 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
     4681 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     4741 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
     4801 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
     4861 acatactcca atactcctga aatagttcat tatggacagc gcagaaagag ctggggagaa
     4921 ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4981 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5041 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5101 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5161 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5221 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     7081 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     7141 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     7201 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     7261 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     7321 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     7381 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     7441 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     7501 cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     7561 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     7621 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     7681 cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag
     7741 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     7801 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aagggggg
//
