LOCUS       sequence-306224-        2733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Source of murine TCR-alpha constant region (TRAC*02) gene followed 
            by P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2733)
  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 2733)
  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: CaP2AIII_pK18
FEATURES             Location/Qualifiers
     source          1..2733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    61..82
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(97..102,103..120,121..127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    135..151
                     /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)."
     primer_bind     140..162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             171..1010
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSGQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTME
                     SGTFITDKTVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEK
                     SFETDMNLNFQNLSVMGLRILLLKVAGFNLLMTLRLWSGSGATNFSLLKQAGDVEENPG
                     PMSCRLLLYVSLCLVETALMWSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGECDLFNKAAVPSSQRNALPVLQPINQF"
     CDS             606..662
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(776..793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(776..792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(785..807)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1011..1826)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLA*A*SRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1734..1753
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1818..1837
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2149..2733,1..4)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2638..2657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
       61 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      121 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacacat atgaccatga
      181 ttacgaattc tggccagaac ccagaacctg ctgtgtacca gttaaaagat cctcggtctc
      241 aggacagcac cctctgcctg ttcaccgact ttgactccca aatcaatgtg ccgaaaacca
      301 tggaatctgg aacgttcatc actgacaaaa ctgtgctgga catgaaagct atggattcca
      361 agagcaatgg ggccattgcc tggagcaacc agacaagctt cacctgccaa gatatcttca
      421 aagagaccaa cgccacctac cccagttcag acgttccctg tgatgccacg ttgaccgaga
      481 aaagctttga aacagatatg aacctaaact ttcaaaacct gtcagttatg ggactccgaa
      541 tcctcctgct gaaagtagcg ggatttaacc tgctcatgac gctgaggctg tggtccggct
      601 ctggagccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa aaccccggac
      661 ctatgagctg caggcttctc ctctatgttt ccctatgtct tgtggaaaca gcactcatgt
      721 ggagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcactgg
      781 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      841 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      901 cccaacagtt gcgcagcctg aatggcgaat gcgatttatt caacaaagcc gccgtcccgt
      961 caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac
     1021 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1081 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1141 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1201 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1261 gagaatggca agagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1321 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgacta cgcctaagcg
     1381 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1441 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1501 acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1561 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1621 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1681 gcatcgggct tcccatacaa ccgatagatt gtcgcacctg attgcccgac attatcgcga
     1741 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cttcgagcaa
     1801 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     1861 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     1921 gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     1981 catcttcccg acaacgcaga ccgttccgtg cgtggctccc tcactttctg gctggatgat
     2041 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctctcgac
     2101 ggagttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2281 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2701 tttgtgatgc tcgtcagggg ggcggagcct atg
//
