LOCUS       sequence-357363-        4499 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 4499)
  AUTHORS   Altman lab
  TITLE     Altman lab bacterial expression plasmids
  JOURNAL   (unpublished)
REFERENCE   2  (bases 1 to 4499)
  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            "(unpublished)"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: pJA5 H-2K(b)
FEATURES             Location/Qualifiers
     source          1..4499
                     /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             1016..1033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1086..1104)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1100..1147
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1184..1639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1271..1290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1481..1502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1666..1770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1771..1839,1840..2631)
                     /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(1989..2008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2802..3390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3291..3310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3544..3561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3576..3718
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3644..3663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3804..3826
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3820..4011)
                     /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 atgggaccac attcgctgag
      121 gtatttcgtc accgccgtgt cccggcccgg cctcggggag ccccggtaca tggaagtcgg
      181 ctacgtggac gacacggagt tcgtgcgctt cgacagcgac gcggagaatc cgagatatga
      241 gccgcgggcg cggtggatgg agcaggaggg gcccgagtat tgggagcggg agacacagaa
      301 agccaagggc aatgagcaga gtttccgagt ggacctgagg accctgctcg gctactacaa
      361 ccagagcaag ggcggctctc acactattca ggtgatctct ggctgtgaag tggggtccga
      421 cgggcgactc ctccgcgggt accagcagta cgcctacgac ggctgcgatt acatcgccct
      481 gaacgaagac ctgaaaacgt ggacggcggc ggacatggcg gcgctgatca ccaaacacaa
      541 gtgggagcag gctggtgaag cagagagact cagggcctac ctggagggca cgtgcgtgga
      601 gtggctccgc agatacctga agaacgggaa cgcgacgctg ctgcgcacag attccccaaa
      661 ggcccatgtg acccatcaca gcagacctga agataaagtc accctgaggt gctgggccct
      721 gggcttctac cctgctgaca tcaccctgac ctggcagttg aatggggagg agctgatcca
      781 ggacatggag cttgtggaga ccaggcctgc aggggatgga accttccaga agtgggcatc
      841 tgtggtggtg cctcttggga aggagcagta ttacacatgc catgtgtacc atcaggggct
      901 gcctgagccc ctcaccctga gatgggagcc tcctccatcc ggatccctgc atcatattct
      961 ggatgcacag aaaatggtgt ggaatcatcg ttaaaagctt gcggccgcac tcgagcacca
     1021 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1081 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1141 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1201 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1261 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1321 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1381 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1441 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1501 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1561 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1621 atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2281 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3601 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     3661 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3721 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3781 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3841 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3901 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3961 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     4021 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     4081 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     4141 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     4201 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     4261 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     4321 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     4381 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     4441 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccg
//
