LOCUS       sequence-462098-        6746 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 6746)
  TITLE     Nathans Lab plasmids 2024
REFERENCE   2  (bases 1 to 6746)
  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-990
FEATURES             Location/Qualifiers
     source          1..6746
                     /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      4052..4061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4065..4199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4102..4121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4156..4175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4268..4625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4268..4459
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4291..4311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4645..4662)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4645..4661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4654..4676)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4874..5329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4961..4980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5171..5192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5346..5365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5465..5487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5525..5543)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5611..5715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5716..5784,5785..6576)
                     /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(5934..5953)
                     /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 acccacgcgt ccggccccat ccccagggat aaccccggag
     1981 cccagagcct caagaaaaaa ttcgttgggc aggggagaga ggtcgcggca gcggcatggc
     2041 aaggttctgg agggccgacc tggccgcagc aggagttatg ttactttgtc actttttaac
     2101 agaccggttc cagttcgccc acggggagcc tggacaccat accaatgatt ggatttatga
     2161 agttacaaac gcttttcctt ggaatgaaga gggggtagaa gtggactctc aagcatacaa
     2221 ccacaggtgg aaaagaaatg tggacccttt taaggcagta gacacaaaca gagccagcat
     2281 gggccaagcc tctccagagt ccaaagggtt cactgacctg ctactggatg acggacagga
     2341 caataacacc cagatagagg aggacacgga tcacaattac tacatttctc ggatatatgg
     2401 tccagcggat tctgccagcc gggatctgtg ggttaacata gaccaaatgg aaaaagacaa
     2461 agtgaagatt cacgggatac tttccaacac tcatcggcaa gctgcaagag tgaatctgtc
     2521 cttcgatttt ccattttatg gtcattttct aaatgaagtc actgtggcaa ctgggggttt
     2581 catatatact ggagaagttg tacatcgaat gctcacagct acacagtata tagctccttt
     2641 aatggcaaat tttgatccca gtgtatccag aaattcaact gtcagatatt ttgataatgg
     2701 cacagctctt gttgtccagt gggaccatgt ccacctgcag gataattaca acctgggaag
     2761 cttcacattc caggccacac tcctcatgga cgggcgcatc atctttggat acaaagaaat
     2821 ccctgtcttg gtcacacaga taagttctac caaccatcca gtgaaagtcg ggttgtctga
     2881 tgcatttgtc gtggtccaca ggatccagca aatacccaat gttcgaagaa gaacaattta
     2941 tgaatatcac cgagtagaac tacaaatgtc caaaattacc aacatctcag ctgtggagat
     3001 gactccactt cccacatgtc tccagttcaa tggttgtggc ccttgtgtgt cctcgcagat
     3061 tggtttcaac tgcagttggt gcagcaaact tcaaagatgc tccagtggat ttgatcgcca
     3121 tcggcaggac tgggtggaca gtggatgccc ggaagaggta cagtcaaaag agaagatgtg
     3181 tgagaagaca gagccaggag agacatctca aactaccacg acctcccaca cgaccaccat
     3241 gcaattcagg gtcctgacca ccaccaggag agctgtgaca tctcagatgc ctaccagcct
     3301 gcctacagaa gatgacacga agatagccct acatctcaaa gacagtggag cctccacaga
     3361 tgacagtgca gctgagaaga aaggaggaac cctccatgca ggcctcattg ttggaattct
     3421 catcttggtc ctcattatag cagcggccat tctggtgaca gtgtatatgt atcaccatcc
     3481 aacatcagca gccagcatct tcttcattga gagacgccca agcagatggc cagcaatgaa
     3541 gtttcgaaga ggctcaggac accctgccta tgcagaagtt gaaccagttg gagagaaaga
     3601 aggttttatt gtatcagagc agtgctaaaa ttttaggaca gagcagcacc agtactggct
     3661 tacaggtgtt aagactaaaa ctttgcttat gcatttaaga caaacagaca cacaacccac
     3721 aaccacacac aaaggagccc taaactgctg tagacagaag ggcgacgaga tttctggaca
     3781 agcccagccc aggaacattg aaaggaaaac tcagacttgt acaagacacc atgtacaatg
     3841 attaaagaat tccctagtgg aatgacatcc atggttcaca aggaacatct ccggtggact
     3901 tgccaggagt gtgacgagat gacgatgctt ttggtttagg tgcagggttg caaagaaatc
     3961 aaggaaaaaa aatatgacaa taaataaagc tttagttcac aaggaaaaaa aaaaaaaaaa
     4021 aaaagggcgg ccgccaccgc ggtggagctt ggccgccatg gcccaacttg tttattgcag
     4081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     4201 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     4261 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4681 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4741 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4801 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4861 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4921 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4981 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5041 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5101 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5161 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     5221 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5281 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5341 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5401 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5461 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5521 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5581 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5641 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5701 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5761 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5821 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5881 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5941 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6001 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6061 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6121 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6181 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6241 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6301 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6361 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6421 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6481 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6541 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6601 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6661 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6721 cccgtagaaa agatcaaagg atcttc
//
