LOCUS       sequence-461150-        6352 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 6352)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 6352)
  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-2016
FEATURES             Location/Qualifiers
     source          1..6352
                     /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    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1020..1399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1400..1603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1554..1574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1600..1624
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1834..1852
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1834..1851
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1865..1884
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3462..3485
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3507..3551
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(3623..3639)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3624..3640)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     regulatory      3658..3667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3671..3805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3708..3727)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3762..3781
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3874..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3874..4065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4251..4268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4251..4267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4260..4282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4480..4935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4567..4586)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4777..4798
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4952..4971)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5071..5093
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5131..5149)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5217..5321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5322..5390,5391..6182)
                     /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(5540..5559)
                     /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 tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg
     1021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     1081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     1501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     1561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     1621 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     1681 cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt
     1741 aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa
     1801 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaataacatc
     1861 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc
     1921 ccaccgcggt ggcggccgcc accatgtccc gtgtgcgccg ccttctgctt ggatacctgt
     1981 tcccagccct cctgttgcat gggctgggag agggctctgc cctccttcat ccagacagca
     2041 gatcgcaccc tcggtcctta gagaaaagcg cctggagggc tttcaaggag tcacagtgtc
     2101 atcacatgct gaagcatctc cacaacggtg cgcggatcac agtgcagatg cccccgacca
     2161 tcgagggcca ctgggtgtcc acaggctgtg aagtaaggtc gggtccggag ttcatgacaa
     2221 ggtcttacag gttctacaac aataatacct tcaaggccta ccagttttac tatggcagca
     2281 accgctgtac aaaccccacc tacaccctca tcatccgagg caagatccgg cttcgccagg
     2341 cgtcctggat catccgtggg ggcaccgaag ctgactacca gcttcacggc gtccaagtca
     2401 tctgccacac agaggcagtc gctgaacagc tcagccgact ggtgaaccga acttgcccag
     2461 gcttcctggc tcctggtggt ccctgggtac aggacgtagc ctatgacctg tggcaggagg
     2521 agagtaacca cgagtgcacc aaggctgtga actttgccat gcacgagctg cagctcatcc
     2581 gtgtggagaa gcagtatccc caccacagcc tggaccacct ggtggaggag ctcttcctgg
     2641 gcgacatcca cacggacgct acccagaggg tgttctaccg gccgtccagt taccagccgc
     2701 ccctgcagaa tgccaagaac cacaaccatg cgtgcatagc ctgccgcatc attttccggt
     2761 cagatgaaca ccaccctccc atactgcccc ccaaggctga cctgaccatt ggcctccacg
     2821 gggaatgggt gagccagcgc tgcgaggtac gccccgaggt cctcttcctc acccgccact
     2881 tcatcttcca cgacaacaac aacacctggg aagggcatta ctaccactac tcagaccctg
     2941 tctgcaagca ccccacattc accatctacg ctcgaggccg ctacagccgc ggtgtgctct
     3001 catctaaggt catgggtggc acggagtttg tgttcaaagt gaatcacatg aaggttactc
     3061 ccatggacgc agccacagcc tccctcctca atgtcttcag tgggaatgag tgtggggctg
     3121 agggctcctg gcaggtgggt atccaacagg atgtgacaca taccaatggc tgcgtggctc
     3181 tgggcatcaa actacctcac acagaatatg agatcttcaa aatggagcaa gacacccgag
     3241 gccgctacct gctgttcaat ggccagaggc ccagcgatgg ctccagccca gacagaccag
     3301 agaagagagc cacatcctac cagatgccct tggtccagtg tgcctcttcc tcaccaagag
     3361 ctgaagagtt gttggaagac agtcaaggtc atctgtatgg cagggcagca gggaggacag
     3421 ctgggtccgg tggttctggt ggcgcgcctg gttctggtgg tcatcatcac caccatcacc
     3481 atcactccgc gcctggttct ggtggtggac tgaacgacat cttcgaggct caaaagatcg
     3541 agtggcacga gggttccgcg cctggtggtt ccaagacaga gaccagccaa gtggcgcctg
     3601 cctaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc gagcttggcc
     3661 gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3781 caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat
     3841 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc
     4261 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4321 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4381 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     4441 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4501 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4561 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4621 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4681 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4741 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4801 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4861 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4921 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4981 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5041 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5101 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5161 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5221 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5281 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5701 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5881 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
