LOCUS       sequence-395025-        7463 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 7463)
  TITLE     Jeff Miner Lab Plasmids
REFERENCE   2  (bases 1 to 7463)
  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_mini-NtA-Fs1-mSA-Myc-His
FEATURES             Location/Qualifiers
     source          1..7463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    24..307
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        498..877
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        878..1081
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1032..1052
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1217..1349
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1332..1351
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1393..1412
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1393..1411
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1512..1521
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2256..2597
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             2598..2618
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2643..2672
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2682..2711
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2721..2750
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2760..2789
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2799..2828
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2856..2873
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2910..3498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2963..2983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3381..3392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3586..3604)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3641..3762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3652..3671)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3706..3725
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4007..4364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4007..4198
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4030..4050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4215..4350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4395..5432
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5411..5419
                     /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    5456..5504
                     /note="synthetic polyadenylation signal"
     promoter        5660..5764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5765..5833,5834..6625)
                     /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(5983..6002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6783..7371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6863..6882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
       61 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
      121 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
      181 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
      241 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
      301 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
      361 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
      421 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      481 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      541 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      601 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      661 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      721 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      781 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      841 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      901 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      961 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
     1021 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1081 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
     1141 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
     1201 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1261 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1321 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1381 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgccg aactcaccta
     1441 tctccccaac acctaataac attcaatcac tctttccact aaccacctat ctacatcacc
     1501 acccaccagc cgccaccatg gtccgcccgc ggctgagctt cccggccccg ctgctgccgc
     1561 tgctgctgct cctggcggcg gccgcgccag ccgtgccggg cgccagcggg acgtgcccgg
     1621 agcgcgcctt ggagcggcgc gaggaggagg cgaacgtggt gctcaccggc acagtggagg
     1681 agatcctcaa cgtggacccg gtgcagcaca cctactcctg caaggttcga gtgtggcggt
     1741 acttgaaagg caaagatgtg gtggcccaag agagcctgct ggatggtggc aacaaggtgg
     1801 taattggtgg ctttggagac cccctcatct gcgacaacca ggtgtccacc ggggacacta
     1861 gaatcttctt tgtgaaccca gcaccaccct acctgtggcc agcccacaag aatgagttga
     1921 tgctcaactc cagccttatg cggatcactt tgcggaacct ggaagaggtg gagttctgtg
     1981 tagaagacaa acctgggatc cacttcacag cagcgccttc tatgcctcct gatgtgtgca
     2041 ggggaatgtt atgtggcttt ggtgctgtgt gtgaacctag tgttgaggat ccaggccggg
     2101 cctcctgtgt gtgcaagaag aatgtctgcc ctgctatggt agctcctgtg tgtggctcag
     2161 atgcttccac ctatagcaac gagtgtgagc tacagcgtgc acagtgcaac cagcaacggc
     2221 gcatccgcct actccgccaa gggccatgtg gctctgcgga agcgggtatc accggcacgt
     2281 ggtacaacca gtctggttct accttcaccg ttaccgcggg tgcggacggt aacctgaccg
     2341 gtcagtacga aaaccgtgcg cagggcactg gttgccagaa ctctccgtac accctgaccg
     2401 gtcgttacaa cggtaccaaa ctggaatggc gtgttgaatg gaacaactct accgaaaact
     2461 gccactctcg taccgaatgg cgtggtcagt accagggtgg tgcggaagcg cgtatcaaca
     2521 cccagtggaa cctgacctac gaaggtggtt ctggtccggc gaccgaacag ggtcaggaca
     2581 ccttcaccaa agttaaagag gatctgtact ttcagagcat cgatggaagt ggtaaagcta
     2641 tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag ctcatttctg
     2701 aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg aatgaaatgg
     2761 agcaaaagct catttctgaa gaggacttga atgaaatgga gcaaaagctc atttctgaag
     2821 aggacttgaa tgaaatggag agcttgggcg acctccatca tcaccatcac cactagggga
     2881 tatcagctga attctagaaa ttcatcgaca atcaacctct ggattacaaa atttgtgaaa
     2941 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3001 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3061 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3121 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3181 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3241 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3301 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3361 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3421 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3481 tttgggccgc ctccccgccc aggttccttg ggccgcgact ctagggcatg caagctgatc
     3541 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     3601 tagcataacc ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt
     3661 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3721 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3781 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     3841 tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     3901 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac
     3961 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     4021 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4201 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4261 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4321 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc gattcttctg
     4381 acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa aaatttctca
     4441 tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa gagagccgag
     4501 ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg
     4561 gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta cccatccccg
     4621 aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt agacgcgccc
     4681 aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg
     4741 ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc ttcggcccat
     4801 tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc gccattgctg
     4861 atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc agcgtagctc
     4921 aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc
     4981 atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg
     5041 actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc ttcttctggc
     5101 ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg catcccgagc
     5161 tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat cagctctacc
     5221 agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca
     5281 tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg agcgcagccg
     5341 tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg cccagtacac
     5401 gaccgcgcgc taaggaggta ggtcgagttt agactctagc catcacgatg gccgcaataa
     5461 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     5521 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     5581 gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg
     5641 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5701 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5761 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5821 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5881 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5941 ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt
     6001 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6061 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6121 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6181 tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6481 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6601 tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta
     6661 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6721 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6781 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6841 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6901 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6961 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7021 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7081 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7141 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7201 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7261 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7321 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     7381 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7441 gtcatgagat tatcaaaaag gat
//
