LOCUS       sequence-425189-        4945 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses murine b2m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  TITLE     pET-3a - mb2m
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4945)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET-3a_mb2m
FEATURES             Location/Qualifiers
     source          1..4945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             35..226
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(220..242)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    328..467
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     383..402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(482..499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(653..1241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(733..752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1412..2203,2204..2272))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2035..2054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2273..2377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2445..2463
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2490..2495,2496..2512,2513..2518)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2884..2931)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2927..2945
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2997..3029)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             3347..3369
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3400..3419)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3401..3419)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3503..3522)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
       61 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
      121 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
      181 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      241 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      301 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
      361 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc
      421 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
      481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1081 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1321 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1381 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1441 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1501 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1561 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1621 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1681 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     1741 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1801 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1861 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1921 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1981 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     2041 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2101 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2161 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2221 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2341 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2401 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2461 ggccctttcg tcttcaagaa ttctcatgtt tgacagctta tcatcgataa gctttaatgc
     2521 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat
     2581 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg
     2641 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat
     2701 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca
     2761 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc
     2821 gactacgcga tcatggcgac cacacccgtc ctgtggatat ccggatatag ttcctccttt
     2881 cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc
     2941 agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg gatccgaccc
     3001 atttgctgtc caccagtcat gctagccatg gtcatcacat gtctcgatcc cagtagacgg
     3061 tcttgggctc ggccatactg tcatgcttaa ctctgcaggc gtatgtatca gtctcagtgg
     3121 gggtgaattc agtgtgagcc aggatataga aagaccagtc cttgctgaag gacatatctg
     3181 acatctctac tttaggaatt tttttcccgt tcttcagcat ttggatttca atgtgaggcg
     3241 ggtggaactg tgttacgtag cagttcagta tgttcggctt cccattctcc ggtgggtggc
     3301 gtgagtatac ttgaatttga ggggttttct ggatgcccat ggtatatctc cttcttaaag
     3361 ttaaacaaaa ttatttctag agggaaaccg ttgtggtctc cctatagtga gtcgtattaa
     3421 tttcgcggga tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc
     3481 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
     3541 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     3601 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
     3661 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
     3721 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     3781 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     3841 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     3901 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc
     3961 accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc
     4021 tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc
     4081 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
     4141 gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt
     4201 ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca
     4261 tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg
     4321 agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc
     4381 caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc
     4441 agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg
     4501 ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     4561 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     4621 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     4681 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     4741 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     4801 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     4861 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     4921 ctcgtttcat cggtatcatt acccc
//
