LOCUS       sequence-355754-        4495 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 4495)
  AUTHORS   Altman lab
  TITLE     Altman lab bacterial expression plasmids
  JOURNAL   (unpublished)
REFERENCE   2  (bases 1 to 4495)
  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            "(unpublished)"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..4495
                     /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        1665..1769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1770..1838,1839..2630)
                     /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(1988..2007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2801..3389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3290..3309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3543..3560
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3575..3714
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3640..3659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3800..3822
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3816..4007)
                     /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 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1741 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1801 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1861 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1921 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1981 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2041 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2101 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2161 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2221 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2281 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2341 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2401 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2461 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2521 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2581 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2641 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2761 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2821 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2881 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2941 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3001 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3061 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3421 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3481 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3541 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3601 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3661 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3721 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3781 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3841 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3901 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3961 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4021 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4081 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4141 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4201 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4261 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4321 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4381 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4441 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccg
//
