LOCUS       sequence-395024-        7790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NtA-Fs1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  TITLE     Jeff Miner Lab Plasmids
REFERENCE   2  (bases 1 to 7790)
  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: pCI_E01_NtA-Fs1-mIgG1Fc-Myc-His-WPRE
FEATURES             Location/Qualifiers
     source          1..7790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    30..313
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        504..883
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        884..1087
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1038..1058
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1223..1355
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1338..1357
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1399..1418
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1399..1417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1518..1527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2931..2951
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2976..3005
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3015..3044
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3054..3083
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3093..3122
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3132..3161
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3189..3206
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3243..3831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3296..3316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3714..3725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3919..3937)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3974..4095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3985..4004)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4039..4058
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4340..4697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4340..4531
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4363..4383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4548..4683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4728..5765
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5744..5752
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5789..5837
                     /note="synthetic polyadenylation signal"
     promoter        5993..6097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6098..6166,6167..6958)
                     /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(6316..6335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7116..7704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7196..7215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc
       61 cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat
      121 gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt
      181 gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg
      241 aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc
      301 tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg
      361 agatcttcaa tattggccat tagccatatt attcattggt tatatagcat aaatcaatat
      421 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
      481 atgtccaata tgaccgccat gttggcattg attattgact agttattaat agtaatcaat
      541 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      601 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      661 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      721 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
      781 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
      841 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      901 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      961 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     1021 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     1081 cagagctggt ttagtgaacc gtcagatcac tagaagcttt attgcggtag tttatcacag
     1141 ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa gctgcagaag
     1201 ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
     1261 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     1321 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag
     1381 ctcttaaggc tagagtatta atacgactca ctatagggct agcaaagcga tcgccgaact
     1441 cacctatctc cccaacacct aataacattc aatcactctt tccactaacc acctatctac
     1501 atcaccaccc accagccgcc accatggtcc gcccgcggct gagcttcccg gccccgctgc
     1561 tgccgctgct gctgctcctg gcggcggccg cgccagccgt gccgggcgcc agcgggacgt
     1621 gcccggagcg cgccttggag cggcgcgagg aggaggcgaa cgtggtgctc accggcacag
     1681 tggaggagat cctcaacgtg gacccggtgc agcacaccta ctcctgcaag gttcgagtgt
     1741 ggcggtactt gaaaggcaaa gatgtggtgg cccaagagag cctgctggat ggtggcaaca
     1801 aggtggtaat tggtggcttt ggagaccccc tcatctgcga caaccaggtg tccaccgggg
     1861 acactagaat cttctttgtg aacccagcac caccctacct gtggccagcc cacaagaatg
     1921 agttgatgct caactccagc cttatgcgga tcactttgcg gaacctggaa gaggtggagt
     1981 tctgtgtaga agacaaacct gggatccact tcacagcagc gccttctatg cctcctgatg
     2041 tgtgcagggg aatgttatgt ggctttggtg ctgtgtgtga acctagtgtt gaggatccag
     2101 gccgggcctc ctgtgtgtgc aagaagaatg tctgccctgc tatggtagct cctgtgtgtg
     2161 gctcagatgc ttccacctat agcaacgagt gtgagctaca gcgtgcacag tgcaaccagc
     2221 aacggcgcat ccgcctactc cgccaagggc catgtggctc tggttgtaag ccttgcatat
     2281 gtacagtccc agaagtatca tctgtcttca tcttcccccc aaagcccaag gatgtgctca
     2341 tgattactct gactcctaag gtcacgtgtg ttgtggtaga catcagcaag gatgatcccg
     2401 aggtccagtt cagctggttt gtagatgatg tggaggtgca cacagctcag acgcaacccc
     2461 gggaggagca gttcaacagc actttccgct cagtcagtga acttcccatc atgcaccagg
     2521 actggctcaa tggcaaggag ttcaaatgca gggtcaacag tgcagctttc cctgccccca
     2581 tcgagaaaac catctccaaa accaaaggca gaccgaaggc tccacaggtg tacaccattc
     2641 cacctcccaa ggagcagatg gccaaggata aagtcagtct gacctgcatg ataacagact
     2701 tcttccctga agacattact gtggagtggc agtggaatgg gcagccagcg gagaactaca
     2761 agaacactca gcccatcatg gacacagatg gctcttactt cgtctacagc aagctcaatg
     2821 tgcagaagag caactgggag gcaggaaata ctttcacctg ctctgtgtta catgagggcc
     2881 tgcacaacca ccatactgag aagagcctct cccactctcc tggtaaagga gaggatctgt
     2941 actttcagag catcgatgga agtggtaaag ctatggagca aaagctcatt tctgaagagg
     3001 acttgaatga aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagcaaa
     3061 agctcatttc tgaagaggac ttgaatgaaa tggagcaaaa gctcatttct gaagaggact
     3121 tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagagcttgg
     3181 gcgacctcca tcatcaccat caccactagg ggatatcagc tgaattctag aaattcatcg
     3241 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3301 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3361 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3421 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3481 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3541 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3601 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     3661 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3721 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3781 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cccaggttcc
     3841 ttgggccgcg actctagggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
     3901 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
     3961 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4021 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4081 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4141 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
     4201 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4261 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     4321 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4501 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4561 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4621 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4681 gcctaggctt ttgcaaaaag ctcgattctt ctgacactag cgccaccatg aagaagcccg
     4741 aactcaccgc taccagcgtt gaaaaatttc tcatcgagaa gttcgacagt gtgagcgacc
     4801 tgatgcagtt gtcggagggc gaagagagcc gagccttcag cttcgatgtc ggcggacgcg
     4861 gctatgtact gcgggtgaat agctgcgctg atggcttcta caaagaccgc tacgtgtacc
     4921 gccacttcgc cagcgctgca ctacccatcc ccgaagtgtt ggacatcggc gagttcagcg
     4981 agagcctgac atactgcatc agtagacgcg cccaaggcgt tactctccaa gacctccccg
     5041 aaacagagct gcctgctgtg ttacagcctg tcgccgaagc tatggatgct attgccgccg
     5101 ccgacctcag tcaaaccagc ggcttcggcc cattcgggcc ccaaggcatc ggccagtaca
     5161 caacctggcg ggatttcatt tgcgccattg ctgatcccca tgtctaccac tggcagaccg
     5221 tgatggacga caccgtgtcc gccagcgtag ctcaagccct ggacgaactg atgctgtggg
     5281 ccgaagactg tcccgaggtg cgccacctcg tccatgccga cttcggcagc aacaacgtcc
     5341 tgaccgacaa cggccgcatc accgccgtaa tcgactggtc cgaagctatg ttcggggaca
     5401 gtcagtacga ggtggccaac atcttcttct ggcggccctg gctggcttgc atggagcagc
     5461 agactcgcta cttcgagcgc cggcatcccg agctggccgg cagccctcgt ctgcgagcct
     5521 acatgctgcg catcggcctg gatcagctct accagagcct cgtggacggc aacttcgacg
     5581 atgctgcctg ggctcaaggc cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg
     5641 gtcgcacaca aatcgctcgc cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc
     5701 tggccgacag cggcaaccgc cggcccagta cacgaccgcg cgctaaggag gtaggtcgag
     5761 tttagactct agccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5821 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     5881 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5941 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6301 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6481 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     6541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     6961 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     7021 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7081 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7141 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7201 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7261 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7321 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7381 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7441 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7501 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7561 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7681 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     7741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
//
