LOCUS       sequence-355758-        4005 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 4005)
  AUTHORS   Altman lab
  TITLE     Altman lab bacterial expression plasmids
  JOURNAL   (unpublished)
REFERENCE   2  (bases 1 to 4005)
  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            "(unpublished)"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..4005
                     /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(1794..1813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1913..1935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1973..1991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2059..2163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2164..2232,2233..3024)
                     /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     complement(2382..2401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3195..3783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3684..3703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3937..3954
                     /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 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1321 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1381 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1441 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1501 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1561 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1621 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1681 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcct gatgcggtat
     1741 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     1801 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     1861 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1921 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     1981 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3961 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccg
//
