LOCUS       sequence-357372-        4007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of soluble MHC protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4007)
  AUTHORS   Altman lab
  TITLE     Altman lab bacterial expression plasmids
  JOURNAL   (unpublished)
REFERENCE   2  (bases 1 to 4007)
  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: 
            "(unpublished)"
COMMENT     Sequence Label: pJA54 HLA-A*03:01
FEATURES             Location/Qualifiers
     source          1..4007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             91..113
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1036..1054)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1037..1054)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1112..1130)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1126..1173
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1272..1727)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1409..1430)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1621..1640
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1796..1815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1915..1937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1975..1993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2061..2165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2166..2234,2235..3026)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3197..3785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3686..3705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3939..3956
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 attcattaat gcagcccggg agatctcgat cccgcgaaat taatacgact cactataggg
       61 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      121 atggctagcg gttctcattc tatgagatat ttcttcacat ccgtgtcccg gcccggccgc
      181 ggggagcccc gcttcatcgc cgtgggctac gtggacgaca cgcagttcgt gcggttcgac
      241 agcgacgccg cgagccagag gatggagccg cgggcgccgt ggatagagca ggaggggccg
      301 gagtattggg accaggagac acggaatgtg aaggcccagt cacagactga ccgagtggac
      361 ctggggaccc tgcgcggcta ctacaaccag agcgaggccg gttctcacac catccagata
      421 atgtatggct gcgacgtggg gtcggacggg cgcttcctcc gcgggtaccg gcaggacgcc
      481 tacgacggca aggattacat cgccctgaac gaggacctgc gctcttggac cgcggcggac
      541 atggcggctc agatcaccaa gcgcaagtgg gaggcggccc atgaggcgga gcagttgaga
      601 gcctacctgg atggcacgtg cgtggagtgg ctccgcagat acctggagaa cgggaaggag
      661 acgctgcagc gcacggaccc ccccaagaca catatgaccc accaccccat ctctgaccat
      721 gaggccaccc tgaggtgctg ggccctgggc ttctaccctg cggagatcac actgacctgg
      781 cagcgggatg gggaggacca gacccaggac acggagctcg tggagaccag gcctgcaggg
      841 gatggaacct tccagaagtg ggcggctgtg gtggtgcctt ctggagagga gcagagatac
      901 acctgccatg tgcagcatga gggtctgccc aagcccctca ccctgagatg ggagctgtct
      961 tcccagcccg gatccctgca tcatattctg gatgcacaga aaatggtgtg gaatcatcgt
     1021 taaaagcttg agtattctat agtgtcacct aaatcccagc ttgatccggc tgctaacaaa
     1081 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     1141 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa
     1201 cctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1261 atggcgaatg gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atatttgtta
     1321 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1381 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1441 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1501 accatcaccc aaatcaagtt ttttgcggtc gaggtgccgt aaagctctaa atcggaaccc
     1561 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1621 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1681 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc ctgatgcggt
     1741 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     1801 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     1861 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     1921 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     1981 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2101 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt catacactat
     2401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg ggcgcggtat tctcagaatg
     2461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2701 cgatgcctgt agcaatgcca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3781 cgaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3901 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3961 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccg
//
