LOCUS       sequence-360103-        7533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a multi-neurotrophin made from the Human NT-3 gene. The 
            secreted protein that can bind only all 3 TRK receptors and not 
            p75NTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7533)
  AUTHORS   Urfer R, Tsoulfas P, Soppet D, Escandon E, Parada LF, Presta LG
  TITLE     The binding epitopes of neurotrophin-3 to its receptors trkC and 
            gp75 and the design of a multifunctional human neurotrophin.
  JOURNAL   EMBO J. 1994 Dec 15. 13(24):5896-909.
  PUBMED    7529173
REFERENCE   2  (bases 1 to 7533)
  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: "EMBO J. 
            1994 Dec 15. 13(24):5896-909."
COMMENT     Sequence Label: pLV-MNTS-1-p75
FEATURES             Location/Qualifiers
     source          1..7533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(136..155)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      949..1537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1438..1457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1691..1708
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1825..1846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1861..1866,1867..1884,1885..1891)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1899..1915
                     /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)."
     primer_bind     1904..1926
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1923..1939
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1923..1939
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1958..1978
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1960..1978
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2006..2232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2233..2413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2460..2585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3078..3311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3496..3540
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    3807..3923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3946..4249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4250..4453
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4404..4424
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4450..4474
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    5382..5970
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5435..5455)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5853..5864)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6057..6290
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6362..6483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6399..6418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6453..6472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6523..6658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6678..6697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6679..6697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6707..6724)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6707..6723)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6716..6738)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6865..7320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6952..6971)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7162..7183
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7346..7450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7451..7519,7520..7533,1..778)
                     /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"
ORIGIN
        1 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
       61 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      121 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      181 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      241 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      301 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      361 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
      421 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
      481 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
      541 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
      601 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
      661 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
      721 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
      781 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
      841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
      901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1561 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1621 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     1681 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     1741 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1801 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     1861 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     1921 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     1981 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
     2041 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
     2101 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
     2161 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
     2221 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     2281 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     2341 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     2401 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     2461 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     2521 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     2581 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     2641 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     2701 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     2761 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     2821 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     2881 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     2941 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     3001 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     3061 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     3121 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     3181 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     3241 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     3301 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     3361 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     3421 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     3481 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     3541 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     3601 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     3661 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     3721 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     3781 aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt
     3841 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     3901 caaattacaa aattcaaaat tttatcgata agcttgggag ttccgcgtta cataacttac
     3961 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4021 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4081 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4141 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4201 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     4261 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     4321 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     4381 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     4441 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     4501 tgacctccat agaagacacc gcccgggcgt acgggatccg ccaccatgag catcctgttc
     4561 tacgtgatct tcctggccta cctgcggggc atccagggca acaacatgga ccagcggagc
     4621 ctgcccgagg acagcctgaa ctccctgatc atcaagctga tccaggccga catcctgaag
     4681 aacaagctga gcaagcagat ggtggacgtg aaagagaact accagagcac cctgcccaag
     4741 gccgaggccc ccagagagcc tgaaagaggc ggccctgcca agagcgcctt ccagcccgtg
     4801 atcgccatgg acaccgagct gctcagacag cagcggcggt acaacagccc cagagtgctg
     4861 ctgagcgaca gcacccccct ggaacccccc cctctgtacc tgatggaaga ttacgtgggc
     4921 agccccgtgg tggccaaccg gaccagcaga cggaagagaa gcagcagcca ccccatcttc
     4981 caccggggcg aggccagcgt ctgtgcctct gagagcctgt gggtcacaga caagagcagc
     5041 gccatcgaca tccggggcca ccaggtcaca gtgctgggcg agatcaagac cggcaacagc
     5101 cccgtgaagc agtacttcta cgagacacgg tgcaaagagg ccagacccgt caagaacggc
     5161 tgccggggca tcgacgacaa gcactggaac agccagtgca agaccagcca gacctacgtg
     5221 cgggccctga ccagcgagaa caacaagctg gtcggatggc ggtggatcag aatcgacacc
     5281 agctgcgtgt gcgccctgag cgccgccatc ggcagaacat gagctagctc tagacctgca
     5341 gggtcgactc cggatgatca gggccctgta cagatatcga caatcaacct ctggattaca
     5401 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5461 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5521 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5581 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5641 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5701 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5761 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5821 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5881 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5941 gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc
     6001 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6061 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     6121 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6181 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6241 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6301 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6361 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6421 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6481 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     6541 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6601 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6661 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     6721 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6781 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6841 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6901 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6961 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     7021 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     7081 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     7141 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     7201 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     7261 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     7321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7501 ttgccttcct gtttttgctc acccagaaac gct
//
