LOCUS       sequence-461167-        6110 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 6110)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 6110)
  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-1093
FEATURES             Location/Qualifiers
     source          1..6110
                     /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      1938..1947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3258..3274
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3258..3274
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3279..3305
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3312..3338
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3345..3371
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3381..3397)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3382..3398)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     regulatory      3416..3425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3429..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3466..3485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3520..3539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3632..3989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3632..3823
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3655..3675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3840..3975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4009..4026)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4009..4025)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4018..4040)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4238..4693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4325..4344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4535..4556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4710..4729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4829..4851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4889..4907)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4975..5079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5080..5148,5149..5940)
                     /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(5298..5317)
                     /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 accatggggc tctccatgga ccgcagccca tacgcccgca
     1981 cgggggacca gcagcgcggc tgctggtact acctgcgcta tttcttcctc ttcgtgtcgc
     2041 tcattcagtt cctcatcatc ctgggcctgg tcctcttcat gatctatggc aatgtgcacg
     2101 ccaccactga gtccagcctg cgcgccacgg agatccgcgc cgacagcctg tacagccagg
     2161 tggttggact atcggcctca caggctaacc tgagcaaaca gctgaacatc agcttgcttg
     2221 tcaaggaaac agtcatgcag caactgttga ctacgcgacg tgagatggag cgcatcaacg
     2281 ccagcttccg ccagtgccaa ggcgacctga tcacctacat aaactataat cgcttcatcg
     2341 ccgctatcat cctgagcgag aagcagtgcc aggaacagct gaaggaggtc aacaagacct
     2401 gcgaagcttt actcttcaag ctgggagaaa aagttaagac actggagatg gaggtggcca
     2461 aggagaaggc agtgtgctcc aaggacaagg agagcctgct ggcaggaaag cggcagacgg
     2521 aagagcagct ggaggcctgt ggcaaagcac gcgaaaggca acagcaggag cagcaggtga
     2581 ccgaggagaa cctgcgcaag gtgcagtccc tgtgcatccc actggaccag gagaagttcc
     2641 aggcggacgt gctgagtgcg tggcgggact cgctgatcta ccgtaccttg gaaaccctgc
     2701 cctaccacta ccagctgatg ccggagtacg cgtccctccg tcggacctgc gagagcctgc
     2761 ccgggatcat gaccaccaag atagaagagc tggcccgcgg gctgcgcgcc ggcattgagc
     2821 gcgtgacccg tgagaatgca gaactgcggc gccagaagtt ggagctagag cgcgcggcac
     2881 aggcggcgca ggaggcgcgg gcgcgcgccg ggaccgaggc gcaggcgcga gaaacccagc
     2941 tacgggcgga gtgcgcgcgc cagacgcagc tggctctgga ggagaaggcg gcgctgcgcg
     3001 cgcagcggga caacctagag cgcgagctgg aggcgcgtaa gcgggagctg gagcaactgc
     3061 gcaccgaggt ggatgtgcgc attagcgcgc tggacacctg cgtcaaggcc aagtcgctgc
     3121 cagccgtccc gccgagagtc tcaggcccac ccccgaaccc tccacccatt gatccagcta
     3181 gcctggagga attcaagaaa aggatcctgg agtctcagcg gctccctgta gtcaaccctg
     3241 ctgcccaacc cagcggttct agaactagtg gatccggcta tccctatgac gtccccgatt
     3301 acgcgggtgg ttacccatac gatgtgccag actatgccgg ggggtaccct tatgacgtgc
     3361 ctgactacgc ttaagcttat cgataccgtc gacctcgagg gggggcccga gcttggccgc
     3421 catggcccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3541 atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa
     3601 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     3661 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3721 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3841 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3901 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3961 gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt
     4021 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4081 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4141 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     4201 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4261 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4321 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4381 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4441 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4501 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4561 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     4621 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4681 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4741 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4801 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4861 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4921 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5041 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5101 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5161 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5221 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5281 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5341 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5401 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5461 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5521 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5581 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5641 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5701 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5761 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5821 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5881 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5941 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6001 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6061 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
