LOCUS       sequence-320949-        5776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MSCV backbone with murine TCR-alpha signal peptide and murine 
            TCR-beta2 constant region (TRBC2) for expression of TCR genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5776)
  AUTHORS   Anderson AM, Landry LG, Alkanani AA, Pyle L, Powers AC, Atkinson MA,
            Mathews CE, Roep BO, Michels AW, Nakayama M
  TITLE     Human islet T cells are highly reactive to preproinsulin in type 1 
            diabetes.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Oct 12;118(41). pii: 2107208118. doi:
            10.1073/pnas.2107208118.
  PUBMED    34611019
REFERENCE   2  (bases 1 to 5776)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-10-12"; volume: "118(41). pii: 
            2107208118. doi"; pages: " 10.1073/pnas.2107208118"
COMMENT     Sequence Label: MBC2-null
FEATURES             Location/Qualifiers
     source          1..5776
                     /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"
     LTR             2171..2685
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2847..2863
                     /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(2871..2877,2878..2895,2896..2901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2916..2937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3054..3071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3225..3813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3305..3324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3984..4775,4776..4844))
                     /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     4607..4626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4845..4949)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5017..5035
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5073..5095)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5195..5214
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5408..5430
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5558..5577
                     /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 tcacgcgtca ccatgaaaac atatgctcct
     1441 acattattca tgtttctatg gctgcagctg gatgggatga gtcgacgtac cagttagatc
     1501 tgagaaatgt gactccaccc aaggtctcct tgtttgagcc atcaaaagca gagattgcaa
     1561 acaaacaaaa ggctaccctc gtgtgcttgg ccaggggctt cttccctgac cacgtggagc
     1621 tgagctggtg ggtgaatggc aaggaggtcc acagtggggt cagcacggac cctcaggcct
     1681 acaaggagag caattatagc tactgcctga gcagccgcct gagggtctct gctaccttct
     1741 ggcacaatcc tcgaaaccac ttccgctgcc aagtgcagtt ccatgggctt tcagaggagg
     1801 acaagtggcc agagggctca cccaaacctg tcacacagaa catcagtgca gaggcctggg
     1861 gccgagcaga ctgtggaatc acttcagcat cctatcatca gggggttctg tctgcaacca
     1921 tcctctatga gatcctactg gggaaggcca ccctatatgc tgtgctggtc agtggcctgg
     1981 tgctgatggc catggtcaag aaaaaaaatt cctgaggatc ccaattgctc gacctgcagg
     2041 catgcaagct tcaggtagcc ggctaacgtt aacaaccggt acctctagaa ctatagctag
     2101 catgcgcaaa tttaaagcgc tgatatcgat aaaataaaag attttattta gtctccagaa
     2161 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     2221 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     2281 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     2341 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     2401 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     2461 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     2521 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     2581 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     2641 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     2701 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     2761 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     2821 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     2881 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2941 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3001 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4981 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5041 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5101 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5161 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5221 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5281 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5341 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5401 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     5461 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5521 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5581 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     5641 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     5701 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     5761 tccagaaaaa gggggg
//
