LOCUS       sequence-331325-        5787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the b1 domain from the human Neuropilin 1 
            receptor (NRP1). With an N-terminal 6His tag and thrombin cleavage 
            site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5787)
  AUTHORS   Daly JL, Simonetti B, Klein K, Chen KE, Williamson MK, Anton-Plagaro
            C, Shoemark DK, Simon-Gracia L, Bauer M, Hollandi R, Greber UF, 
            Horvath P, Sessions RB, Helenius A, Hiscox JA, Teesalu T, Matthews 
            DA, Davidson AD, Collins BM, Cullen PJ, Yamauchi Y
  TITLE     Neuropilin-1 is a host factor for SARS-CoV-2 infection.
  JOURNAL   Science. 2020 Oct 20. pii: science.abd3072. doi: 
            10.1126/science.abd3072.
  PUBMED    33082294
REFERENCE   2  (bases 1 to 5787)
  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: "Science. 
            2020 Oct 20. pii: science.abd3072. doi: 10.1126/science.abd3072."
COMMENT     Sequence Label: pET28a(+) NRP1 b1 (273-427)
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     604..623
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(643..720)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     926..945
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1029..1048
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1029..1047
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1048..1072
                     /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)."
     RBS             1087..1109
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1128..1145
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1155..1172
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1680..1697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1750..1768)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1764..1811
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1848..2303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1935..1954)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2145..2166
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2395..3210)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2395..3210)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3118..3137
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3202..3221
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3332..3920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3821..3840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4074..4091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4106..4245
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4171..4190)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4331..4353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4347..4538)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5347..5787,1..642))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
       61 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
      121 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
      181 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
      241 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
      301 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
      361 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
      421 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
      481 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
      541 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
      601 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
      661 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
      721 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
      781 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      841 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      901 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      961 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     1021 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     1081 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
     1141 atcacagcag cggcctggtg ccgcgcggca gccatatgga aaacctgtat cagggcggcc
     1201 acatgtttaa atgtatggaa gcactgggca tggaaagtgg tgaaattcat agtgatcaga
     1261 ttaccgccag cagtcagtat agcaccaatt ggagtgccga acgtagccgc ctgaattatc
     1321 cggaaaatgg ttggaccccg ggtgaagata gctatcgcga atggattcag gttgatctgg
     1381 gcctgctgcg ttttgttacc gccgttggta cccagggtgc cattagtaaa gaaaccaaaa
     1441 agaaatacta cgtgaagacc tataagattg atgttagcag caatggtgaa gattggatta
     1501 ccattaagga aggtaataag ccggttctgt ttcagggcaa taccaatccg accgatgttg
     1561 tggttgccgt ttttccgaaa ccgctgatta cccgttttgt tcgtattaag ccggccacct
     1621 gggaaaccgg tattagtatg cgctttgaag tttatggttg caaaattacc taactcgagc
     1681 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1741 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1801 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
     1861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1921 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1981 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2041 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2101 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2161 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2221 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2281 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     2341 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     2401 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2461 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2521 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2581 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2641 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     2701 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2761 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2821 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2881 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     2941 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     3001 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     3061 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     3121 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     3181 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     3241 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4081 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4141 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4201 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4261 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4321 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4381 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     4441 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     4501 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     4561 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4621 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4681 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4741 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4801 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4861 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4921 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4981 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     5041 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     5101 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     5161 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     5221 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     5281 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     5341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5461 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5521 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5581 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5641 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5701 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5761 catggcactc cagtcgcctt cccgttc
//
