LOCUS       sequence-473591-        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression in bacterial cells for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Dankert JF, Pagan JK, Starostina NG, Kipreos ET, Pagano M
  TITLE     FEM1 proteins are ancient regulators of SLBP degradation.
  JOURNAL   Cell Cycle. 2017 Mar 19;16(6):556-564. doi: 
            10.1080/15384101.2017.1284715. Epub 2017 Jan 24.
  PUBMED    28118078
REFERENCE   2  (bases 1 to 6218)
  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"; doi: 
            "10.1080/15384101.2017.1284715"; journalName: "Cell Cycle"; date: 
            "2017-03-19- 19"; volume: "16"; issue: "6"; pages: "556-564"
COMMENT     Sequence Label: pUC18 SLBP
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     618..637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(856..960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1035..1054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1226..1605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1606..1809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1760..1780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1854..1873
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1854..1872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1889..1918
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2762..2780)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2763..2780)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2800..2817)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2806..3030
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3076..3504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3373..3394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3513..3533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3518..3847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3698..3833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3760..3779
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3914..4708
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3968..3987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4578..4597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4882..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4973..4992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5052..5068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5052..5068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5065..5087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5076..5092
                     /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)."
     promoter        complement(join(5100..5106,5107..5124,5125..5130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5145..5166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5283..5300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5454..6042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5534..5553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6213..6218,1..786,787..855))
                     /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"
ORIGIN
        1 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
       61 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      121 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      301 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
      721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
      781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      841 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
      901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
      961 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     1021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     1081 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     1141 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     1201 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     1261 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1321 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1381 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1441 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1501 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1561 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1621 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1681 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1741 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1801 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     1861 actcactata gggagaccca agcttatgga gcagaagctc atctcagagg aggacctggg
     1921 atctgcctgc cgcccgcgaa gcccgccgag gcatcagagc cgctgcgacg gtgacgccag
     1981 cccgccgtcc cccgcgcgat ggagcctggg acggaagcgc agagccgacg gcaggcgctg
     2041 gaggcccgaa gacgccgagg aggcagagca ccgcggcgcc gagcgcagac ccgagagctt
     2101 taccactcct gaaggcccta aaccccgttc cagatgctct gactgggcaa gtgcagttga
     2161 agaagatgaa atgaggacca gagttaacaa agaaatggca agatataaaa ggaaactcct
     2221 catcaatgac tttggaagag agagaaaatc atcatcagga agttctgatt caaaggagtc
     2281 tatgtctact gtgccggctg actttgagac agatgaaagt gtcctaatga ggagacagaa
     2341 gcagatcaac tatgggaaga acacaattgc ctacgatcgt tatattaaag aagtcccaag
     2401 acaccttcga caacctggca ttcatcccaa gacccctaat aaatttaaga agtatagtcg
     2461 acgttcatgg gaccagcaaa tcaaactctg gaaggtggct ctgcattttt gggatcctcc
     2521 agcggaagaa ggatgtgatt tgcaagaaat acaccctgta gaccttgaat ctgcagaaag
     2581 cagctccgag ccccagacca gctctcagga tgactttgat gtgtactctg gcacacccac
     2641 caaggtgaga cacatggaca gtcaagtgga ggatgagttt gatttggaag cttgtttaac
     2701 tgaacccttg agagacttct cagccatgag ctaactcgag catgcatcta gagggcccta
     2761 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     2821 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2881 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2941 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3001 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3061 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3121 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3181 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3241 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3301 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3361 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3421 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3481 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3781 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3841 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3901 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3961 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4021 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4081 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4141 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4201 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4261 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4321 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4381 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4441 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4501 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4561 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4621 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4681 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4741 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4801 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5641 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5881 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6181 taaagtatat atgagtaaac ttggtctgac agttacca
//
