LOCUS       sequence-456206-        7576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NXPE1 protein residues 60-547 in mammalian cells (HEK293T 
            or expi293). x6 His tag and IL2 secretion signal included N terminal
            with GGGG linker to protein sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7576)
  TITLE     NXPE1 Protein Producing Plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7576)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NXPE1_pcDNA3.4-TOPO
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     sig_peptide     754..813
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             814..831
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2353..2941
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2406..2426)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2824..2835)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2962..2981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3006..3054
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3256..3684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3343..3362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3553..3574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3693..3713)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3698..4027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3878..4013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3940..3959
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4094..4888
                     /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(4148..4167)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4758..4777
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5064..5185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5101..5120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5155..5174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5234..5250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5234..5250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5247..5269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5258..5274
                     /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(5282..5288,5289..5306,5307..5312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5327..5348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5465..5482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5636..6224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5716..5735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6395..7186,7187..7255))
                     /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     7018..7037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7256..7360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7435..7454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaaccct tggatccgcc accatgtaca gaatgcagct gctgagctgt
      781 atcgccctgt ctctggccct ggtcaccaat tctcaccacc accatcacca cctgattcct
      841 ctgaagcccc tgaccgagac agagctgcgg atcaaagaga tcatcgagaa gctggaccag
      901 cagatccctc ctcggccttt cacacacgtg aacaccacaa caagcgccac acacagcacc
      961 gccacaatcc tgaatcctcg ggatacctac tgcagaggcg accagctgga cattctgctg
     1021 gaagtgcggg atcacctggg acagagaaag cagtacggcg gcgatttcct gagagccaga
     1081 atgagcagtc ctgctctgac agccggcgct agcggcaaag tgatggactt caacaacggc
     1141 acctacctgg tgtctttcac cctgttctgg gaaggccagg tttccctgag cctgctgctg
     1201 attcatcctt ctgaaggcgc cagcgctctg tggcgagcca gaaatcaggg ctacgacaag
     1261 atcatcttca agggcaagtt cgtgaacggc accagccacg tgttcaccga gtgtggcctg
     1321 acactgaaca gcaatgccga gctgtgcgag tacctggacg acagagatca agaggccttc
     1381 tactgcatga agccccagca catgccttgc gaggccctga cctacatgac caccagaaac
     1441 agagaggtgt cctacctgac cgacaaagag aacagcctgt tccaccggtc caaagtgggc
     1501 gtcgagatga tgaaggaccg gaagcacatc gacgtgacca actgcaacaa gcgggaaaag
     1561 atcgaagaga catgccaagt gggaatgaag cctcctgtgc ctggcggcta tacactgcaa
     1621 ggcaagtgga tcaccacgtt ctgcaatcaa gtgcagctcg acaccatcaa gatcaacggc
     1681 tgcctgaagg gcaagctgat ctacctgctg ggcgatagca ccctgcggca gtggatctac
     1741 tacttcccca aggtggtcaa gaccctgaag ttcttcgacc tgcacgagac agggatcttc
     1801 aagaagcacc tcctgctgga cgccgagaga cacacacaga tccagtggaa gaagcacagc
     1861 taccccttcg tgaccttcca gctgtactcc ctgatcgacc acgactacat ccccagagag
     1921 atcgacagac tgagcggcga caagaacacc gccatcgtga tcaccttcgg ccagcacttc
     1981 agaccctttc caatcgacat cttcattcgg agagccatcg gcgtgcagaa ggccatcgag
     2041 agactgtttc tgagaagccc cgccaccaaa gtgatcatca agaccgagaa catccgcgag
     2101 atgcacatcg aaaccgagag attcggcgac tttcacggct acatccacta cctgatcatg
     2161 aaggacatct ttaaggacct gaacgtgggc atcatcgacg cctgggacat gacaatcgcc
     2221 tacggcaccg ataccatcca tcctccagac cacgtgatcg gcaaccagat caacatgttc
     2281 ctgaactaca tctgctgatg actcgagaag ggttcgatcc ctaccggtta gtaatgagtt
     2341 tgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2401 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2461 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2521 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2581 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2641 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2701 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2761 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2821 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2881 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2941 cctggaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     3001 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     3061 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     3121 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     3181 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     3241 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3301 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3361 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3421 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3481 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3541 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3601 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3661 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3901 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3961 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4021 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4081 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4141 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4201 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4261 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4321 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4381 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4441 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4501 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4561 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4621 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4681 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4741 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4801 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4861 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     4921 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4981 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5041 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5101 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5161 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5221 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7261 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7321 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7381 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7441 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7501 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7561 gaagaatctg cttagg
//
