LOCUS       sequence-462382-        7252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7252)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 7252)
  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"
COMMENT     Sequence Label: pPMS-1438
FEATURES             Location/Qualifiers
     source          1..7252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    889..910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(925..930,931..948,949..955)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    963..979
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     968..990
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     987..1003
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     987..1003
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1346..1365
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1364..1419
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1465..1484
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1580..1603)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(2691..2720)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2736..2759)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(2781..2804)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     intron          4411..5427
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4465..4487)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        5707..6086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6117..6221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6222..6290,6291..7082)
                     /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(6440..6459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat cgatccgctg
      841 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      901 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      961 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     1021 tgggctgcag gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc
     1081 aggagaggag gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag
     1141 aaaaaaataa cacaaaacaa aatataaaaa aaatctaacc tcaagtcaag gcttttctat
     1201 ggaataagga atggacagca gggggctgtt tcatatactg atgacctctt tatagccaac
     1261 ctttgttcat ggcagccagc atatgggcat atgttgccaa actctaaacc aaatactcat
     1321 tctgatgttt taaatgattt gccctcccat atgtccttcc gagtgagaga cacaaaaaat
     1381 tccaacacac tattgcaatg aaaataaatt tcctttatta gccagaagtc agatgctcaa
     1441 ggggcttcat gatgtcccca taatttttgg cagagggaaa aagatctcag tggtatttgt
     1501 gagccagggc attggccaca ccagccacca ccttctgata ggcagcctgc acctgaggag
     1561 tgaattatcg ctcgagtcaa tgatggtgat gatggtggtg atggaattca agcatgcccg
     1621 cgggcggccg cacccagcgt cctgcggcag ctgggttctc aaccgctggg ttgggcagca
     1681 gggttgacta cagggagccg ctgagactcc aggatccttt tcttgaattc ctccaggcta
     1741 gctggatcaa tgggtggagg gttcgggggt gggcctgaga ctctcggcgg gacggctggc
     1801 agcgacttgg ccttgacgca ggtgtccagc gcgctaatgc gcacatccac ctcggtgcgc
     1861 agttgctcca gctcccgctt acgcgcctcc agctcgcgct ctaggttgtc ccgctgcgcg
     1921 cgcagcgccg ccttctcctc cagagccagc tgcgtctggc gcgcgcactc cgcccgtagc
     1981 tgggtttctc gcgcctgcgc ctcggtcccg gcgcgcgccc gcgcctcctg cgccgcctgt
     2041 gccgcgcgct ctagctccaa cttctggcgc cgcagttctg cattctcacg ggtcacgcgc
     2101 tcaatgccgg cgcgcagccc gcgggccagc tcttctatct tggtggtcat gatcccgggc
     2161 aggctctcgc aggtccgacg gagggacgcg tactccggca tcagctggta gtggtagggc
     2221 agggtttcca aggtacggta gatcagcgag tcccgccacg cactcagcac gtccgcctgg
     2281 aacttctcct ggtccagtgg gatgcacagg gactgcacct tgcgcaggtt ctcctcggtc
     2341 acctgctgct cctgctgttg cctttcgcgt gctttgccac aggcctccag ctgctcttcc
     2401 gtctgccgct ttcctgccag caggctctcc ttgtccttgg agcacactgc cttctccttg
     2461 gccacctcca tctccagtgt cttaactttt tctcccagct tgaagagtaa agcttcgcag
     2521 gtcttgttga cctccttcag ctgttcctgg cactgcttct cgctcaggat gatagcggcg
     2581 atgaagcgat tatagtttat gtaggtgatc aggtcgcctt ggcactggcg gaagctggcg
     2641 ttgatgcgct ccatctcacg tcgcgtacca cctctagaac cagaaccatt caagtcctct
     2701 tcagaaatga gcttttgctc accactagag gatccgggcc cctggaacag aacttccagg
     2761 gaggagggga tgtcgtagtc gtggtggtgg tggtggtggt ggtggccgct gaagccacag
     2821 ctgccctcca cagagcggca ctgcacgatg cgcaggaatg tctcgacctt gtccatgtcc
     2881 ttcctgaagc agtagagcag cccgtagttc ttgagtagtg cgtcatcgtt gtgtgagttt
     2941 gtgtcgaact tgctgtaggt ctgcttgaag atctgcccag tccgggggct gccatcttcc
     3001 agcctctgca aagtgaagga agagaaggag aggccaagcg cttgggtact gttccctccc
     3061 tttctcattc attcattttc ctccctcccc ttcagggtgt agagaaaggc ctgaggattc
     3121 acgaggggaa atgaagaata aggtgagttc tcttgggtca gggcctgact gctaaaaaga
     3181 gggcagcagt gtttctctaa cacagctctc aaagtcagtg gggctccagg attggggacc
     3241 cctggcgcca ccctcacccc catcagcgtt tggatgcctt cctctaggtc ctttaggagg
     3301 tcatagacgt tgctgtcaga ggcgccgtac accaggctgt tggcgaagac actcctgagg
     3361 aactgcacgg gctccagcca cgactggatg agcagcaggg agatgcggag cagctctagg
     3421 ttctgcaggg gaaggacggg cattggctgt gctgcccggg ggctctgact acaggtctcc
     3481 cccatccccg cctggggaga aggcatccac tcacggattt ctgttgtgtt tcctccctgt
     3541 tggagggtgt cggaatagac tctgagaaac agagggaggt ctgggggttc tgcaggaatg
     3601 aatacttctg ttcctttggg atataggctt cttcctagga gaaggaccgc ccaccaaggt
     3661 ctacgctgga gaccagctcc cattgttact tttctgggaa cctcactcag cgtgtgctca
     3721 tctgcctgca ttttcgcttc gggaaaaacc ctgagctcct tagtctcctc ctcttatttc
     3781 ccagcggggg aaagtcaccc cttcctgcca cccctgatgc gcacccattc cccaagagct
     3841 tacaaactcc tggtaggtgt caaaggccag ctggtgcaga cgatgggcgc ggagcatagc
     3901 gttgtcaaaa agcctggata agggaatggt tgggaaggca ctgccctctt gaagccaggg
     3961 caggcagagc aggccaaaag ccaggagcag ggacgtccgg gagcctgggg agaaaccaga
     4021 gggcaacaga gggagccgga gagcaagagg ccagcactct ccctgctcca ggagctgttt
     4081 gtttttctct ctccatccct ccagggacca ggagctttct gagattggcc aaatactggg
     4141 cttacatggc gatactcaca ttcagaagcc ccaaacctga gggttagtgc ccccgtccca
     4201 tctacaggtc gctgcctctc ccctcaggac acattgtgcc caaagggatt ttaggggcgc
     4261 ttacctgtag ccattgcagc taggtgagct gtccacagga ccctgagtgg ttcggggagt
     4321 tgggccttgg gtaccgaact cggcctgaga cggccgcaat tctttgccaa aatgatgaga
     4381 cagcacaaca accagcacgt tgcccaggag ctgtaggaaa aagaagaagg catgaacatg
     4441 gttagcagag gcgctagagc cgccggtcac acgccagaag ccgaaccccg ccctgccccg
     4501 tcccccccga aggcagccgt ccccccgcgg acagccccga ggctggagag ggagaagggg
     4561 acggcggcgc ggcgacgcac gaaggccctc cccgcccatt tccttcctgc cggcgccgca
     4621 ccgcttcgcc ccgcgcccgc tagagggggt gcggcggcgc ctcccagatt tcggctccgc
     4681 acagatttgg gacaaaggaa gtccctgcgc cctctcgcac gattaccata aaaggcaatg
     4741 gctgcggctc gccgcgcctc gacagccgcc ggcgctccgg gggccgccgc gccccycccc
     4801 cgagcccycc ccggcccgmg gcggccccgc cccgcccggc acccccmcct gccgccaccc
     4861 cccgcccggc acggcgagcc ccgcgccacg ccccgtacgg agccccgcac ccgaagccgg
     4921 gccgtgctca gcaactcggg gaggggggtg cagggggggt tgcagcccga ccgacgcgcc
     4981 cacaccccct gctcaccccc ccacgcacac accccgcacg cagcctttgt tcccctcgca
     5041 gccccccccg caccgcgggg caccgccccc ggccgcgctc ccctcgcgca cactgcggag
     5101 cgcacaaagc cccgcgccgc gcccgcagcg ctcacagccg ccgggcagcg cggagccgca
     5161 cgcggcgctc cccacgcaca cacacacgca cgcacccccc gagccgctcc ccccgcacaa
     5221 agggccctcc cggagcccct caaggctttc acgcagccac agaaaagaaa caagccgtca
     5281 ttaaaccaag cgctaattac agcccggagg agaagggccg tcccgcccgc tcacctgtgg
     5341 gagtaacgcg gtcagtcaga gccggggcgg gcggcgcgag gcggcggcgg agcggggcac
     5401 ggggcgaagg cagcgcgcag cgactcccgc ccgccgcgcg cttcgctttt tatagggccg
     5461 ccgccgccgc cgcctcgcca taaaaggaaa ctttcggagc gcgccgctct gattggctgc
     5521 cgccgcmccy cyccsccycs ccccsccccc cccccccccc cccccccccc cccccggcgc
     5581 gcgccccccc cccccccccg cccccatcgc tgcacaaaat aattaaaaaa taaataaata
     5641 caaaattggg ggtggggagg ggggggagat ggggagagtg aagcagaacg tggggctcac
     5701 ctcgaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     5761 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     5821 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     5881 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     5941 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     6001 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     6061 ttaataacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg
     6121 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6181 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6241 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6301 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6361 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6421 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6481 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6541 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6601 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6661 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6721 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6781 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6841 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6901 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6961 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7021 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7081 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7141 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7201 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
