LOCUS       sequence-462102-        5782 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 5782)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 5782)
  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-2272
FEATURES             Location/Qualifiers
     source          1..5782
                     /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"
     regulatory      3088..3097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3101..3235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3138..3157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3192..3211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3304..3661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3304..3495
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3327..3347)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3512..3647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3681..3698)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3681..3697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3690..3712)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3910..4365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3997..4016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4207..4228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4382..4401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4501..4523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4561..4579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4647..4751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4752..4820,4821..5612)
                     /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(4970..4989)
                     /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 accatgaaga ccctgctgct gaccctggtg gtggtgacaa
     1981 tcgtgtgcct ggacctggga tacaccatcg tgtgccacac caccgccacc agccctatca
     2041 gcgctgtgac ctgccctcct ggcgagaacc tgtgctacag aaagatgtgg tgcgacgcct
     2101 tctgtagcag cagaggcaag gtggtcgagc tgggatgcgc cgctacctgc cctagcaaga
     2161 aaccttacga agaggtgacc tgctgtagca ccgacaagtg caaccctcac cctaagcagc
     2221 gccctggagg ttctggtggt tctggaggag gcgcgcctgg ttctggtggt tctggtacat
     2281 gtcccaagtg cccacctcct gaaaacctgg gtggaccatc tgtcttcatc tttcccccga
     2341 agcccaagga cacgctcatg atctccctga cccctagggt cacatgtgtg gtggtagatg
     2401 tgagccaaga tgagcctgaa gtccagttca catggttcgt ggacaacaaa ccggtcggca
     2461 atgctgagac aaagccccga gtggagcaat acaacacgac attccgcgtg gaaagtgtcc
     2521 tccccatcca gcaccaggac tggctgaggg gcaaggaatt caagtgcaag gtctacaaca
     2581 aagccctgcc agcccccata gagaagacca tctccaaaac caaaggtggg agtgcagtgc
     2641 aggcaggtgg gagattggga ggcactgtgg gggtgaaatg gacaggccct ccatggccag
     2701 ccctccatcc ctggcattga acatgtgctg atctctgtcc cacaggggct ccccgcatgc
     2761 cagatgtgta cacccttccc ccgtcccgag acgagctatc caagagcaaa gtcagtgtga
     2821 cctgcctgat catcaacttc tttcctgccg acatccacgt ggagtgggcc agcaataggg
     2881 ttccagtgag tgagaaggaa tacaagaaca ccccacccat tgaggacgct gacgggtcct
     2941 acttcctcta cagcaagctc actgtggata agagcgcgtg ggatcaggga accgtctaca
     3001 cctgctccgt gatgcatgaa gccctgcaca atcatgtcac tcagaaggcc atctcccgct
     3061 ctccgggtaa atgagggccc gagcttggcc gccatggccc aacttgttta ttgcagctta
     3121 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3181 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa
     3241 ttaattcggc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3301 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3601 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3661 aaagctgtta acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3721 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3781 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     3841 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3901 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4321 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4381 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4561 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5761 tagaaaagat caaaggatct tc
//
