LOCUS       sequence-473272-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FEM1A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  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 7400)
  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: pcDNA3.1alphaFEM1AalphaFLAG
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(137..241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(316..335)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        507..886
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        887..1090
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1041..1061
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1135..1154
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1135..1153
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1207..1216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3220..3243
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3263..3280)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3269..3493
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3539..3967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3626..3645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3836..3857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3976..3996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3981..4310
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4161..4296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4223..4242
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4377..5171
                     /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(4431..4450)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5041..5060
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5345..5466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5382..5401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5436..5455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5515..5531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5515..5531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5528..5550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5539..5555
                     /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(5563..5569,5570..5587,5588..5593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5608..5629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5746..5763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5917..6505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5997..6016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6676..7400,1..67,68..136))
                     /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     7299..7318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
       61 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      241 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
      301 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
      361 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
      421 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
      481 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      541 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      601 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      661 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      721 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      781 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      841 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      901 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      961 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     1021 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1081 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     1141 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc
     1201 ggatccgcca ccatggacct ccgcaccgcc gtgtacaacg ccgcccgtga tggcaagctg
     1261 cagctgctcc agaagctgct cagcggccgg agccgggagg aactggacga gctgacgggc
     1321 gaggtggccg gcgggggaac gccgctactc atcgccgccc gctacggcca cctggacgtg
     1381 gtggagtacc tggtggaccg gtgcggcgcg agcgtggagg ccggtggctc ggtgcacttc
     1441 gatggcgaga ccatcgaggg cgcgccgccg ctgtgggccg cctccgcagc cggccacctg
     1501 gacgtggtgc ggagcctgct gcgccgcggg gcctcggtga accgcaccac gcgcaccaac
     1561 tccacgcctc tccgcgccgc ctgcttcgac ggccacctgg aggtggtgcg ctacctggtc
     1621 ggcgagcacc aggccgacct ggaggtggcc aaccggcacg gccacacgtg cctcatgatc
     1681 tcgtgctaca agggccaccg tgagatcgcc cgctacctgc tggagcaggg cgcccaggtg
     1741 aaccggcgca gcgccaaggg caacacggcc ctgcatgact gcgccgagtc cggcagcctg
     1801 gagatcctgc agctgctgct ggggtgcaag gcccgcatgg aacgtgacgg ctacggcatg
     1861 accccgctgc tcgcggccag cgtgacgggc cacaccaaca tcgtggagta cctcatccag
     1921 gagcagcccg gccaggagca ggtcgcaggg ggagaggctc agcctgggct gccccaagaa
     1981 gacccctcca ccagccaggg gtgtgcgcag cctcaggggg ctccgtgctg cagctcctcc
     2041 ccagaggaac cactgaacgg ggaatcttac gaaagctgct gtcccaccag ccgggaagct
     2101 gccgtggaag ccttggaatt gctgggagct acgtatgtgg ataagaaacg agatctgctt
     2161 ggggccctta aacactggag gcgggccatg gagctgcgtc accagggggg cgagtacctg
     2221 cccaaaccgg agcccccaca gctggtcctg gcctatgact attccaggga ggtcaacacc
     2281 accgaggagc tggaggcgct gatcaccgac ccggatgaga tgcgcatgca ggccctgttg
     2341 atccgggagc gcatcctcgg tccctcgcac ccggacactt cctattacat ccgttacagg
     2401 ggtgccgtgt acgccgactc gggcaatttc gagcgctgca tccgcttgtg gaagtacgcc
     2461 ctggacatgc aacagagcaa cctggagcct ctgagcccca tgaccgccag cagcttcctc
     2521 tccttcgcgg aactcttctc ctacgtgctt caggaccggg ccgccaaagg cagcctgggc
     2581 acccagatcg gctttgcaga cctcatgggg gttctcacca aaggggtccg ggaagtggaa
     2641 cgggccctgc agctgcccag ggagcccgga gactcagccc agttcaccaa ggcgctggcc
     2701 atcatcctcc acctgctcta cctgctggag aaagtggagt gcacccccag ccaggagcac
     2761 ctgaagcacc agaccgtcta ccgcctgctc aagtgcgcgc ccaggggcaa gaacggcttc
     2821 acccctctgc acatggctgt ggacaaggac accacaaacg tgggccgcta tcccgtgggc
     2881 agattcccct ccctgcacgt ggtcaaagtg ctgctcgact gcggggccga cccggacagc
     2941 agggattttg acaacaacac cccgctacac atagcagccc agaacaactg cccggccatc
     3001 atgaatgccc tgatcgaagc aggggcccac atggacgcca ccaatgcctt caagaagacg
     3061 gcctacgagc tgctggacga gaagctgctg gccaggggta ccatgcagcc cttcaactac
     3121 gtgaccctgc agtgccttgc ggcccgggcc ctggataaga acaagatccc ttacaagggc
     3181 ttcatcccgg aagatctgga ggcgttcatc gaactgcacg attacaagga tgacgacgat
     3241 aagtgataaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3301 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3361 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3421 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3481 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3541 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3601 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3661 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3721 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3781 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3841 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3901 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3961 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4081 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4141 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4201 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4261 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4321 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4381 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4441 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4501 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4561 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4621 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4681 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4741 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4801 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4861 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4921 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4981 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5041 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5101 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5161 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5221 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5281 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5341 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5401 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5461 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5521 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5581 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5641 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5701 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7381 accgctgttg agatccagtt
//
