LOCUS       sequence-473271-        7274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FEM1B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7274)
  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 7274)
  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: pcDNA3.1alphaFEM1BalphaFLAG
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(108..176,177..968)
                     /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(326..345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1139..1727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1628..1647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1881..1898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2015..2036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2051..2056,2057..2074,2075..2081)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2089..2105
                     /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     2094..2116
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2113..2129
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2113..2129
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2178..2299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2189..2208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2243..2262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2473..3267)
                     /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(2584..2603)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3194..3213
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3334..3663)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3348..3483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3402..3421)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3648..3668
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3677..4105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3999..4018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4151..4375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4364..4381
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4401..4424)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6302..6311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6364..6383)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6365..6383)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6428..6631)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6457..6477)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6632..7011
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7183..7202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
       61 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
      661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1801 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1861 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1921 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1981 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2041 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2101 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     2161 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2221 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2281 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     2341 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     2401 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     2461 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     2521 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     2581 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     2641 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     2701 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     2761 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     2821 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     2881 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     2941 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     3001 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     3061 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     3121 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     3181 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     3241 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga
     3301 tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa
     3361 gcctcctcac tacttctgga atagctcaga ggcagaggcg gcctcggcct ctgcataaat
     3421 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     3481 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     3541 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     3601 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     3661 cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3721 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4021 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4081 cttaatgcgc cgctacaggg cgcgtgggga taccccctag agccccagct ggttctttcc
     4141 gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
     4201 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     4261 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     4321 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
     4381 gctgatcagc gggtttatca cttatcgtcg tcatccttgt aatcatgaaa tccaacaaac
     4441 tcttcaagag ttctggggat ctggtcttgg tagttaatgt cattagcccg aactgctcgg
     4501 gcagccaggc acttgagact catcttcatt tgagttttaa gcagtatttc agatacccca
     4561 gttgtacttt tgtctagcgg agtcttattc tgtttattcg tcatgtcagt gtgagctccg
     4621 gcttcaacta ggctaatgat gatggagtgc aaggtcaaaa aatcactgat gggcctgttg
     4681 tactgaacga taatatgaag ggcactgttt ccctcattgt ccacggcatt cacctcagca
     4741 ccacagtcca gcaggagctt tgtgacaagt gcatttggaa agctgcagac gtcattggtg
     4801 tggaaatcat caactggagt attggaattg acagccagat gcagcaaggt gaaaccttca
     4861 cgagttctgg gatcaaggtg aatcaggttg tagatctgct tgttaatttt gcactgatct
     4921 tcttcgctgc actgtgtttt ggtagagatg cacactaaat acagaaaggt atagagatta
     4981 cattcataat tgtccatagc attgtggaca tcagcatctg aaatattttt cactctgttc
     5041 atactttgtt ctatttccaa aacactgcat ctcaaaacac attctatgtc tggggccttc
     5101 acagtttcat tcaaatgtat catttgtgag aaaacttgag caaatcgaag aagatccttg
     5161 tgggtgttcc tgttaccttt ttgtctgagg tgcagggcat gaagccacaa cttgatacac
     5221 tgctcaaatt ccatattatc cgcataaaca gctcctctgt aaatgatggg atgagaaaca
     5281 tcaatattgt cagcacctaa aatccgttcc cgaactataa ggccttccat atgaagagca
     5341 tctctgtctt gccgaatgga ctccagttcc tgaggatttc tacattcagt tctattccca
     5401 taagcatgga ttggtggaag aacctctttt tcgagaatgt tatcaccatc ttggaacctc
     5461 tctaacatgg ctaaatatag atagtggtat gtctttatga tgtcatagtt ctcacggtca
     5521 tttgcaaagg aggcacccaa gagttccaaa gcttcaatcc gacttcttcg gtcgcaatca
     5581 gcatgagaga gtaacagttc tacgacatca gctttacagc tttcggcagc tactttcaat
     5641 ggcgtcatcc catggccatt cactactata gcagcacgcc attttatcag ctctttcaca
     5701 atatctatgt gcccagcttc agctgcaaag tgcaatgctg tggctccaca atgtgctttg
     5761 gcattgggat cagcacgttg ttctaaaagg tatctgacca catcagtgtg tcccttatat
     5821 gccgcaatca ttaggcaggt gttgtcatat ttgttggcaa tgctgatgtt ggcattattt
     5881 tcaaccaagt atttcacaat gtccagtctg ccatcaaagc atgctgcccg caggggggtt
     5941 gaattagtta ctgtggtatg gttcacgttg gctccatggc tgactagaag tttaacaact
     6001 tcaaaatgtc ctgctccagc tgcacaccaa agagcagtgg caccatcaat gacatacccg
     6061 tcgaagcgga cggtgccagt ctgctgagtc tgcacccggt aatgttctaa gagcaagcgt
     6121 accacctttg cgtgtccatt gcgggctgcg atgatgaggg gcgtggagcg ctgccctccc
     6181 tgctggctga catagccaag cagatagcgg atgtcgcttt cagaccggtt gagaagcaag
     6241 gcggccagag tcagcacctt gccctcgctg gccgccttgt atacatagcc agccaggccc
     6301 tccatggtgg cggatccgag ctcggtacca agcttaagtt taaacgctag ccagcttggg
     6361 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
     6421 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     6481 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     6541 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     6601 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     6661 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     6721 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     6781 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     6841 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     6901 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     6961 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
     7021 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     7081 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     7141 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     7201 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
     7261 ttttcgggga aatg
//
