LOCUS       sequence-360100-        7536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a multi-neurotrophin made from the Human NT-3 gene. The 
            secreted protein can bind all 3 TRK receptors and p75NTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7536)
  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 7536)
  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
FEATURES             Location/Qualifiers
     source          1..7536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(93..112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      906..1494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1395..1414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1648..1665
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1782..1803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1818..1823,1824..1841,1842..1848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1856..1872
                     /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     1861..1883
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1880..1896
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1880..1896
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1915..1935
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1917..1935
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1963..2189
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2190..2370
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2417..2542
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3035..3268
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3453..3497
                     /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    3764..3880
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3903..4206
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4207..4410
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4361..4381
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4407..4431
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    5342..5930
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5395..5415)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5813..5824)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6017..6250
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6322..6443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6359..6378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6413..6432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6483..6618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6638..6657)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6639..6657)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6667..6684)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6667..6683)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6676..6698)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6825..7280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6912..6931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7122..7143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7306..7410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7411..7479,7480..7536,1..735)
                     /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 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
       61 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      121 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      181 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      241 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
      301 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
      361 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
      421 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
      481 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
      541 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
      601 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
      661 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
      721 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
      781 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
      841 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
      901 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      961 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1021 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1081 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1141 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1201 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1261 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1321 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1381 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1441 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1501 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1561 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     1621 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1681 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1741 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     1801 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     1861 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1921 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     1981 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     2041 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     2101 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     2161 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     2221 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     2281 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     2341 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     2401 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     2461 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     2521 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     2581 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     2641 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     2701 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     2761 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     2821 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     2881 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2941 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     3001 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     3061 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     3121 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     3181 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     3241 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     3301 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     3361 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     3421 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     3481 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     3541 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     3601 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     3661 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     3721 gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg
     3781 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     3841 ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt atcgataagc ttgggagttc
     3901 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3961 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4021 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4081 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4141 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4201 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     4261 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4321 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4381 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     4441 cgccatccac gctgttttga cctccataga agacaccgcc cgggcgtacg ggatccgcca
     4501 ccatgagcat cctgttctac gtgatcttcc tggcctacct gcggggcatc cagggcaaca
     4561 acatggacca gcggagcctg cccgaggaca gcctgaactc cctgatcatc aagctgatcc
     4621 aggccgacat cctgaagaac aagctgagca agcagatggt ggacgtgaaa gagaactacc
     4681 agagcaccct gcccaaggcc gaggccccca gagagcctga aagaggcggc cctgccaaga
     4741 gcgccttcca gcctgtgatc gccatggaca ccgagctgct caggcagcag cggcggtaca
     4801 acagccccag agtgctgctg agcgacagca cccccctgga acccccccct ctgtacctga
     4861 tggaagatta cgtgggcagc cccgtggtgg ccaaccggac cagcagacgg aagcggagca
     4921 gcagccaccc catcttccac cggggcgagt acagcgtgtg cgccagcgag agcctgtggg
     4981 tcaccgacaa gagcagcgcc atcgacatcc ggggccacca ggtcacagtg ctgggcgaga
     5041 tcaagaccgg caacagcccc gtgaagcagt acttctacga gacacggtgc aaagaggcca
     5101 gacccgtcaa gaacggctgc cggggcatcg acgacaagca ctggaacagc cagtgcaaga
     5161 ccagccagac ctacgtgcgg gccctgacca gcgagaacaa caagctcgtg ggctggcggt
     5221 ggatcagaat cgacaccagc tgcgtgtgtg ccctgagccg gaagatcggc cggacctgat
     5281 gagctagctc tagacctgca gggtcgactc cggatgatca gggccctgta cagatatcga
     5341 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5401 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5461 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5521 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5581 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5641 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5701 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     5761 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5821 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5881 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg
     5941 agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6001 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     6061 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6121 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6181 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6241 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     6301 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     6361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6421 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     6481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6541 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6601 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     6661 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     6721 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     6781 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     6841 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6901 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6961 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7021 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7081 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7141 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7201 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     7261 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     7321 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     7381 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     7441 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7501 gtaaaagatg ctgaagatca gttgggtgca cgagtg
//
