LOCUS       sequence-357368-        4502 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 4502)
  AUTHORS   Altman lab
  TITLE     Altman lab bacterial expression plasmids
  JOURNAL   (unpublished)
REFERENCE   2  (bases 1 to 4502)
  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            "(unpublished)"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: pJA13 Mamu-A1*002:01
FEATURES             Location/Qualifiers
     source          1..4502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1019..1036
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1089..1107)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1103..1150
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1187..1642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1274..1293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1484..1505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1669..1773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1774..1842,1843..2634)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1992..2011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2805..3393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3294..3313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3547..3564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3579..3721
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3647..3666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3807..3829
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3823..4014)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgggatctc attcaatgag
      121 atatttctac acctccatgt cccggcccgg ccgctgggag ccccgcttca tcgccgtggg
      181 ctacgtggac gacacgcagt tcgtgcggtt cgacagcgac gccgcgagcc agaggatgga
      241 gccgcgggcg ccgtgggtgg agcaggaggg tccagagtat tgggaccggg agacacggaa
      301 catgaaggcc gagacacaga atgccccagt gaacctgcgg aacctgcgcg gctactacaa
      361 ccagagcgag gccgggtctc acaccatcca gaggatgtac ggctgcgacc tggggccgga
      421 cgggcgcctc ctccgcgggt atcaccagtc cgcctacgac ggcaaggatt acatcgccct
      481 gaacgaggac ctgcgctcct ggaccgccgc ggacatggcg gctcagaaca cccagcgcaa
      541 gtgggaggcg gcgggtgagg cggagcagca cagaacctac ctggagggcg agtgcctgga
      601 gtggctccgc agatacctgg agaacgggaa ggagacgctg cagcgcgcgg acccccccaa
      661 gacacatgtg acccaccacc ccgtctctga ccaagaggcc accctgaggt gctgggccct
      721 gggcttctac cctgcggaga tcacactgac ctggcagcgg gatggagagg accaaactca
      781 ggacacggag ctcgtggaga ccaggcctgc aggggacgga accttccaga agtgggcggc
      841 tgtggtggtg ccttctggaa aggagcagag atacacctgt catgtgcagc atgagggtct
      901 gcgtgagccc ctcaccctga gatgggagcc gtcttcccag tccggatccc tgcatcatat
      961 tctggatgca cagaaaatgg tgtggaatca tcgttaaaag cttgcggccg cactcgagca
     1021 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1081 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1141 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     1201 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1261 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1321 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1381 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1441 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1501 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1561 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1621 aaaatattaa cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1861 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1921 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1981 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2041 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2101 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2161 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2221 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2281 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     2341 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2401 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2461 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2521 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2581 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2641 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2701 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2761 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2821 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2881 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2941 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3001 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3061 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3121 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3181 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3241 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3301 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3361 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3421 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3481 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3541 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     3601 acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg
     3661 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3721 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3781 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3841 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3901 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3961 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     4021 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     4081 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     4141 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     4201 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     4261 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     4321 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     4381 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     4441 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     4501 cg
//
