LOCUS       sequence-421637-        8355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express myc-NRP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8355)
  TITLE     Res. Sq.-- NRP1 interacts with endoglin and VEGFR2 to modulate VEGF 
            signaling and endothelial cell sprouting
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8355)
  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            "Unpublished"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: myc-tagged NRP1 (human)
FEATURES             Location/Qualifiers
     source          1..8355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..47)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             991..1020
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3761..3778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3767..3991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4037..4465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4124..4143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4334..4355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4474..4494)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4479..4808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4659..4794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4721..4740
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4857..5882
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4857..5882
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6012..6133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6049..6068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6103..6122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6182..6198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6182..6198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6195..6217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6206..6222
                     /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(6230..6236,6237..6254,6255..6260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6275..6296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6413..6430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6584..7172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6664..6683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7343..8134,8135..8203))
                     /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     7966..7985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8204..8308)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
       61 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      181 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      841 cgaaattaat acgactcact atagggagac ccaagctggc tagcgtttaa acttaagctt
      901 ggtaccgagc tcggatccac catggagagg gggctgccgc tcctctgcgc cgtgctcgcc
      961 ctcgtcctcg ccccggccgg cgcttttcgc gaacaaaaac tcatctcaga agaggatctg
     1021 aacgataaat gtggcgatac tataaaaatt gaaagccccg ggtaccttac atctcctggt
     1081 tatcctcatt cttatcaccc aagtgaaaaa tgcgaatggc tgattcaggc tccggaccca
     1141 taccagagaa ttatgatcaa cttcaaccct cacttcgatt tggaggacag agactgcaag
     1201 tatgactacg tggaagtctt cgatggagaa aatgaaaatg gacattttag gggaaagttc
     1261 tgtggaaaga tagcccctcc tcctgttgtg tcttcagggc catttctttt tatcaaattt
     1321 gtctctgact acgaaacaca tggtgcagga ttttccatac gttatgaaat tttcaagaga
     1381 ggtcctgaat gttcccagaa ctacacaaca cctagtggag tgataaagtc ccccggattc
     1441 cctgaaaaat atcccaacag ccttgaatgc acttatattg tctttgcgcc aaagatgtca
     1501 gagattatcc tggaatttga aagctttgac ctggagcctg actcaaatcc tccagggggg
     1561 atgttctgtc gctacgaccg gctagaaatc tgggatggat tccctgatgt tggccctcac
     1621 attgggcgtt actgtggaca gaaaacacca ggtcgaatcc gatcctcatc gggcattctc
     1681 tccatggttt tttacaccga cagcgcgata gcaaaagaag gtttctcagc aaactacagt
     1741 gtcttgcaga gcagtgtctc agaagatttc aaatgtatgg aagctctggg catggaatca
     1801 ggagaaattc attctgacca gatcacagct tcttcccagt atagcaccaa ctggtctgca
     1861 gagcgctccc gcctgaacta ccctgagaat gggtggactc ccggagagga ttcctaccga
     1921 gagtggatac aggtagactt gggccttctg cgctttgtca cggctgtcgg gacacagggc
     1981 gccatttcaa aagaaaccaa gaagaaatat tatgtcaaga cttacaagat cgacgttagc
     2041 tccaacgggg aagactggat caccataaaa gaaggaaaca aacctgttct ctttcaggga
     2101 aacaccaacc ctacagatgt tgtggttgca gtattcccca aaccactgat aactcgattt
     2161 gtccgaatca agcctgcaac ttgggaaact ggcatatcta tgagatttga agtatatggt
     2221 tgcaagataa cagattatcc ttgctctgga atgttgggta tggtgtctgg acttatttct
     2281 gactcccaga tcacatcatc caaccaaggg gacagaaact ggatgcctga aaacatccgc
     2341 ctggtaacca gtcgctctgg ctgggcactt ccacccgcac ctcattccta catcaatgag
     2401 tggctccaaa tagacctggg ggaggagaag atcgtgaggg gcatcatcat tcagggtggg
     2461 aagcaccgag agaacaaggt gttcatgagg aagttcaaga tcgggtacag caacaacggc
     2521 tcggactgga agatgatcat ggatgacagc aaacgcaagg cgaagtcttt tgagggcaac
     2581 aacaactatg atacacctga gctgcggact tttccagctc tctccacgcg attcatcagg
     2641 atctaccccg agagagccac tcatggcgga ctggggctca gaatggagct gctgggctgt
     2701 gaagtggaag cccctacagc tggaccgacc actcccaacg ggaacttggt ggatgaatgt
     2761 gatgacgacc aggccaactg ccacagtgga acaggtgatg acttccagct cacaggtggc
     2821 accactgtgc tggccacaga aaagcccacg gtcatagaca gcaccataca atcagagttt
     2881 ccaacatatg gttttaactg tgaatttggc tggggctctc acaagacctt ctgccactgg
     2941 gaacatgaca atcacgtgca gctcaagtgg agtgtgttga ccagcaagac gggacccatt
     3001 caggatcaca caggagatgg caacttcatc tattcccaag ctgacgaaaa tcagaagggc
     3061 aaagtggctc gcctggtgag ccctgtggtt tattcccaga actctgccca ctgcatgacc
     3121 ttctggtatc acatgtctgg gtcccacatc ggcacactca gggtcaaact gcgctaccag
     3181 aagccagagg agtacgatca gctggtctgg atggccattg gacaccaagg tgaccactgg
     3241 aaggaagggc gtgtcttgct ccacaagtct ctgaaacttt atcaggtgat tttcgagggc
     3301 gaaatcggaa aaggaaacct tggtgggatt gctgtggatg acattagtat taataaccac
     3361 atttcacaag aagattgtgc aaaaccagca gacctggata aaaagaaccc agaaattaaa
     3421 attgatgaaa cagggagcac gccaggatac gaaggtgaag gagaaggtga caagaacatc
     3481 tccaggaagc caggcaatgt gttgaagacc ttagacccca tcctcatcac catcatagcc
     3541 atgagtgccc tgggggtcct cctgggggct gtctgtgggg tcgtgctgta ctgtgcctgt
     3601 tggcataatg ggatgtcaga aagaaacttg tctgccctgg agaactataa ctttgaactt
     3661 gtggatggtg tgaagttgaa aaaagacaaa ctgaatacac agagtactta ttcggaggca
     3721 tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3781 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3841 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3901 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3961 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4021 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4081 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4141 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4201 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4261 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4321 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4381 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4441 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4501 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4561 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4621 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4681 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4741 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4801 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgatga
     4861 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     4921 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     4981 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     5041 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     5101 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     5161 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     5221 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     5281 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     5341 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     5401 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     5461 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     5521 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     5581 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     5641 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     5701 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     5761 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     5821 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     5881 agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5941 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6001 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6121 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6181 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6241 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6301 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6361 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6421 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6481 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7261 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7321 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7381 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7441 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7501 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7561 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7621 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7681 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7741 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7801 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7861 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7921 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7981 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8041 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8101 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8161 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8221 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8281 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8341 acggatcggg agatc
//
