LOCUS       sequence-461166-        6402 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 6402)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 6402)
  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-989
FEATURES             Location/Qualifiers
     source          1..6402
                     /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     1932..1948
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1933..1949
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3666..3682)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3666..3682)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      3708..3717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3721..3855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3758..3777)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3812..3831
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3924..4281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3924..4115
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3947..3967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4132..4267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4301..4318)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4301..4317)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4310..4332)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4827..4848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5002..5021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5121..5143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5181..5199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5267..5371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5372..5440,5441..6232)
                     /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(5590..5609)
                     /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 cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcg cccttctgca
     1981 gccgccgccg ccgccgcgca gacccggagg tcgtccgcgc ccaccatgcg agctcagctc
     2041 tggttgctgc agttgctgct actccgcggg gccgcgcgcg cgctcagtcc cgcaacaccc
     2101 gcaggtcaca atgaaggtca agactctgca tggactgcca agaggaccag gcaaggctgg
     2161 agtcggagac cccgagagag ccccgcgcag gtgttgaagc caggcaagac ccagctaagc
     2221 caggacttgg gtgggggctc cctggccatc gacacacttc cggacaacag gactcgggtg
     2281 gtggaggaca accataacta ctacgtgtcc cgtgtctacg gccctggtga gaaacgaagc
     2341 caggatctgt gggtggacct ggctgtggcc aaccggagtc atgtgaagat ccacaggatc
     2401 ctctccagtt ctcaccgaca ggcttcaaga gtggtcttgt cctttgattt ccctttctat
     2461 gggcatcctc tgcggcagat caccatagca accggaggct tcatcttcat gggggacatg
     2521 ctccaccgga tgctcacagc tacgcagtat gtggcacccc tgatggccaa cttcaacccc
     2581 ggctactctg acaactccac ggttgcttac tttgacaatg ggaccgtttt tgtggttcag
     2641 tgggatcatg tttacctcca ggaccgggag gacaggggca gcttcacctt ccaggcggcc
     2701 ctacaccgag atggccgcat tgtcttcggc tacaaagaga tccccatggc tgtcctggat
     2761 atcagccctg cccagcaccc tgtcaaggca ggcctgtccg acgctttcat gattctcaat
     2821 tcatccccag aggtgccaga gtctcagaga cggaccattt tcgaatacca ccgtgtggaa
     2881 ctggactcca gcaagatcac caccacctcg gccgtggagt tcaccccgtt gccaacctgc
     2941 ctccagcatc agagttgcga cacctgcgtg tcctcgaatc taaccttcaa ctgcagctgg
     3001 tgccatgtcc tgcagaggtg ttccagtggc tttgaccgat accgccaaga atggctgacc
     3061 tacggctgtg cccaggaggc agaaggcaag acatgcgagg acttccagga tgatagccac
     3121 tactcagcct cccctgacag ctccttcagc ccctttaatg gcgactccac tacctcttcc
     3181 tccctcttca ttgacagcct caccacagaa gatgacacca agttgaatcc ctacgcagaa
     3241 ggagacggcc ttccggacca ctcatctccg aagtccaagg gtcctcccgt gcacctgggc
     3301 accatcgtgg gtatcgtgct tgctgtactc ctggtagcgg ccatcatcct ggctgggatt
     3361 tacatcagtg gccaccctaa ttccaatgct gcactcttct tcatcgagcg gagacctcac
     3421 cactggccag ccatgaagtt ccataaccac cccaaccact ctacctacac cgaggtagag
     3481 ccctccgggc acgagaagga aggcttcgtg gaggccgagc agtgctgaga gagcatcggg
     3541 ccagagacct ggggatggca tgaaagacaa gtcagagcac agagaagtga ttttttttcc
     3601 tggcctcctc ggagctggcc ctgggccagg aaaatagatg aagggcgaat tcctgcagcc
     3661 cgggggatcc actagttcta gagcggccgc caccgcggtg gagcttggcc gccatggccc
     3721 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3781 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3841 tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat aacctctgaa
     3901 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4261 ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc gttttacaac
     4321 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4381 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4441 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4501 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4981 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5161 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5221 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
