LOCUS       sequence-473150-        7244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FEM1C in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7244)
  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 7244)
  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: pcDNA3.1alphaFEM1CalphaFLAG
FEATURES             Location/Qualifiers
     source          1..7244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     268..287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(506..610)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(685..704)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        876..1255
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1256..1459
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1410..1430
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1504..1523
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1504..1522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1576..1585
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3433..3456
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3476..3493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3482..3706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3752..4180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3839..3858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4049..4070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4189..4209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4194..4523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4374..4509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4436..4455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4590..5384
                     /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(4644..4663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5254..5273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5558..5679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5595..5614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5649..5668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5728..5744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5728..5744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5741..5763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5752..5768
                     /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(5776..5782,5783..5800,5801..5806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5821..5842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5959..5976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6130..6718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6210..6229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6889..7244,1..436,437..505))
                     /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 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
       61 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
      121 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
      181 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
      241 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
      301 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
      361 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
      421 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
      481 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
      541 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      601 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
      661 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
      721 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
      781 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      841 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      901 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      961 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1021 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1081 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1141 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1201 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1261 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1321 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1381 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1441 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     1501 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttggt
     1561 accgagctcg gatccgccac catggatcta aagacagcag tatttaacgc agctcgggat
     1621 ggcaaactcc ggcttctcac caaattgttg gcaagcaaat ccaaagagga ggtttcctcc
     1681 ttgatctctg aaaaaacaaa tggggccacg ccactcttga tggccgccag gtatgggcac
     1741 cttgacatgg tggaattcct cctagagcaa tgcagtgcct ccatagaagt tgggggctcc
     1801 gtcaattttg atggcgaaac cattgagggg gctccccctt tatgggccgc ttctgcagca
     1861 ggacatctga aggtggtcca gtccttgtta aatcatggag catctgtcaa caacacgact
     1921 ttaaccaatt caactcctct tcgagctgcg tgtttcgatg gccatttgga aatagtgaag
     1981 taccttgtag aacacaaagc tgatttggaa gtgtcaaacc gacatgggca tacgtgcttg
     2041 atgatttcat gttacaaagg acataaagag attgctcagt atttacttga aaagggggca
     2101 gatgttaata gaaaaagtgt caaaggtaat actgcattgc atgattgtgc agaatctgga
     2161 agtttggaca tcatgaagat gcttcttatg tattgtgcca agatggaaaa ggatggttat
     2221 ggaatgactc cccttctctc agcaagtgtg actggtcaca caaatattgt ggattttctg
     2281 acacaccatg cacagaccag caagacagaa cgtattaatg ctctagagct tctgggagct
     2341 acatttgtag acaaaaaaag agatctgctt ggggctttga aatactggaa aaaggcaatg
     2401 aacatgaggt acagtgatag gactaatatt attagtaaac cagtgccaca gacactaata
     2461 atggcttatg attatgccaa ggaagtgaac agtgcagaag agctagaagg tcttattgct
     2521 gatcctgatg agatgagaat gcaggcacta ttaatcagag aacgtattct tggtccttct
     2581 catcctgata cctcttacta tattagatat agaggcgctg tctatgcaga ctctggaaat
     2641 ttcaaacgat gcatcaacct atggaagtat gctttggata tgcagcagag caatttggat
     2701 cctttaagcc caatgaccgc cagcagctta ttatcttttg cagaactatt ctcctttatg
     2761 ctacaggata gggctaaagg cctgctgggt actactgtta catttgatga tcttatgggc
     2821 atactttgca aaagcgtcct tgaaatagag cgagctatca aacaaactca gtgtccagct
     2881 gacccattac agttaaataa ggccctttct attattttgc acttaatttg cttgttagag
     2941 aaagttcctt gtactctaga acaagaccat ttcaaaaagc agactatata caggtttctt
     3001 aagctgcatc caaggggaaa gaataacttc agccctcttc atctggctgt ggacaagaat
     3061 actacatgtg tagggcggta ccctgtttgt aaatttccat ctctacaagt tactgcaata
     3121 ctgatagaat gtggtgctga tgtgaacgtc agagactcgg atgacaacag tcccctgcat
     3181 atcgctgctc ttaacaacca tccagacatc atgaatctcc ttattaaatc aggtgcacat
     3241 tttgatgcca caaacttgca caaacaaact gctagtgact tgctggatga gaaggaaata
     3301 gctaaaaatt taatccagcc tataaatcat accacattgc agtgtcttgc tgctcgtgtc
     3361 atagtgaatc atagaatata ttataaaggg catatcccag aaaagctaga gacttttgtt
     3421 tcccttcata gagattacaa ggatgacgac gataagtgat aaacccgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3721 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3961 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4021 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4081 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4141 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4201 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4561 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4621 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4681 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4741 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4801 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4861 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4921 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4981 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5041 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5101 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5161 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5221 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5281 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5341 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5401 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5461 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5521 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5701 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gctt
//
