LOCUS       sequence-461160-        6651 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 6651)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 6651)
  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-1791
FEATURES             Location/Qualifiers
     source          1..6651
                     /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"
     primer_bind     1940..1956
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1940..1956
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3922..3938)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3923..3939)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     regulatory      3957..3966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3970..4104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4007..4026)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4061..4080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4173..4530
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4173..4364
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4196..4216)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4381..4516
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4550..4567)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4550..4566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4559..4581)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4779..5234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4866..4885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5076..5097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5251..5270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5370..5392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5430..5448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5516..5620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5621..5689,5690..6481)
                     /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(5839..5858)
                     /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 ggcggccgct ctagaactag tggatccccc gggctgcagg aattcgccct
     1981 tcggccactg tacctctgag ctgtgcacgc cgcggccggg gcgggcctcg ggactggggc
     2041 tgggagccaa ggggccgggg cgggacgcgg agaggctggg ctgcggttcg gagtcccgcg
     2101 cgcgcgctcg gagggggaca gagacggact acagcgaggc ccggaggagc cccgagatgt
     2161 cccgtgtgcg ccgccttctg cttggatacc tgttcccagc cctcctgttg catgggctgg
     2221 gagagggctc tgccctcctt catccagaca gcagatcgca ccctcggtcc ttagagaaaa
     2281 gcgcctggag ggctttcaag gagtcacagt gtcatcacat gctgaagcat ctccacaacg
     2341 gtgcgcggat cacagtgcag atgcccccga ccatcgaggg ccactgggtg tccacaggct
     2401 gtgaagtaag gtcgggtccg gagttcatga caaggtctta caggttctac aacaataata
     2461 ccttcaaggc ctaccagttt tactatggca gcaaccgctg tacaaacccc acctacaccc
     2521 tcatcatccg aggcaagatc cggcttcgcc aggcgtcctg gatcatccgt gggggcaccg
     2581 aagctgacta ccagcttcac ggcgtccaag tcatctgcca cacagaggca gtcgctgaac
     2641 agctcagccg actggtgaac cgaacttgcc caggcttcct ggctcctggt ggtccctggg
     2701 tacaggacgt agcctatgac ctgtggcagg aggagagtaa ccacgagtgc accaaggctg
     2761 tgaactttgc catgcacgag ctgcagctca tccgtgtgga gaagcagtat ccccaccaca
     2821 gcctggacca cctggtggag gagctcttcc tgggcgacat ccacacggac gctacccaga
     2881 gggtgttcta ccggccgtcc agttaccagc cgcccctgca gaatgccaag aaccacaacc
     2941 atgcgtgcat agcctgccgc atcattttcc ggtcagatga acaccaccct cccatactgc
     3001 cccccaaggc tgacctgacc attggcctcc acggggaatg ggtgagccag cgctgcgagg
     3061 tacgccccga ggtcctcttc ctcacccgcc acttcatctt ccacgacaac aacaacacct
     3121 gggaagggca ttactaccac tactcagacc ctgtctgcaa gcaccccaca ttcaccatct
     3181 acgctcgagg ccgctacagc cgcggtgtgc tctcatctaa ggtcatgggt ggcacggagt
     3241 ttgtgttcaa agtgaatcac atgaaggtta ctcccatgga cgcagccaca gcctccctcc
     3301 tcaatgtctt cagtgggaat gagtgtgggg ctgagggctc ctggcaggtg ggtatccaac
     3361 aggatgtgac acataccaat ggctgcgtgg ctctgggcat caaactacct cacacagaat
     3421 atgagatctt caaaatggag caagacaccc gaggccgcta cctgctgttc aatggccaga
     3481 ggcccagcga tggctccagc ccagacagac cagagaagag agccacatcc taccagatgc
     3541 ccttggtcca gtgtgcctct tcctcaccaa gagctgaaga gttgttggaa gacagtcaag
     3601 gtcatctgta tggcagggca gcagggagga cagctgggtc cctgttgctt cctgcctttg
     3661 tcagcctttg gactctccca cattggcgca tcctcagata gaagacattt cctgaaccaa
     3721 gactctttac cgtgtatgac tttccttcac acagagaaag gacattgatt cttttgatgc
     3781 acttgaatgc cttgagacct gccgttgcct ctcctctccc cacctcttcc agcctctgcg
     3841 ccatgagcag tgtgtttgaa gtttcagctt tgaatgtctt cccgcccttt tgtccaaggg
     3901 cgaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg agcttggccg
     3961 ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4021 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4081 aatgtatctt atcatgtctg gatcgggaat taattcggcg cagcaccatg gcctgaaata
     4141 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     4201 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4381 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4441 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4501 ggcttttttg gaggcctagg cttttgcaaa aagctgttaa cagcttggca ctggccgtcg
     4561 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4621 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4681 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     4741 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4801 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4861 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4921 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4981 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5041 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5101 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     5161 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5221 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5281 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5341 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5401 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5461 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5521 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5581 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5641 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5701 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5761 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5821 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5881 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5941 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6001 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6061 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6121 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6181 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6241 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6301 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6361 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6421 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6481 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
