LOCUS       sequence-360108-        7395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To expres Human GDNF with optimized codon usage for better 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  TITLE     Neurotrophin related plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7395)
  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            "Unpublished"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: pLV-HuGDNF
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(135..154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      948..1536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1437..1456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1690..1707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1824..1845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1860..1865,1866..1883,1884..1890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1898..1914
                     /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     1903..1925
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1922..1938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1922..1938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1957..1977
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1959..1977
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2005..2231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2232..2412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2459..2584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3077..3310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3495..3539
                     /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    3806..3922
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3945..4248
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4249..4452
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4403..4423
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4449..4473
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    5243..5831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5296..5316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5714..5725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5918..6151
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6223..6344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6260..6279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6314..6333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6384..6519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6539..6558)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6540..6558)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6568..6585)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6568..6584)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6577..6599)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6726..7181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6813..6832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7023..7044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7207..7311
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7312..7380,7381..7395,1..777)
                     /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 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
       61 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
      181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
      661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
      781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     1741 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1801 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1861 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1921 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     1981 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     2041 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     2101 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     2161 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     2221 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     2281 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     2341 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     2401 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     2461 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     2521 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     2581 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     2641 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     2701 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     2761 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     2821 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     2881 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     2941 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     3001 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     3061 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     3121 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     3181 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     3241 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     3301 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     3361 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     3421 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     3481 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     3541 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     3601 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     3661 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     3721 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     3781 acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg
     3841 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     3901 aaattacaaa attcaaaatt ttatcgataa gcttgggagt tccgcgttac ataacttacg
     3961 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4021 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4081 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     4141 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     4201 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     4261 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4321 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4381 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     4441 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     4501 gacctccata gaagacaccg cccgggcgta cgggatccgc caccatgaag ctgtgggacg
     4561 tggtggccgt gtgcctggtg ctgctgcaca ccgccagcgc ctttccactg cctgccggca
     4621 agaggcctcc tgaggctcct gccgaggaca gaagcctggg cagaagaagg gcccccttcg
     4681 ccctgagcag cgacagcaac atgcccgagg actaccccga ccagttcgac gacgtgatgg
     4741 acttcatcca ggccaccatc aagcggctga agcggagccc cgacaagcag atggccgtgc
     4801 tgcccagaag agagcggaac agacaggccg ctgccgccaa ccccgagaac agcagaggca
     4861 agggcagacg gggccagcgg ggcaagaaca gaggctgtgt gctgaccgcc atccacctga
     4921 acgtgaccga cctgggcctg ggctacgaga caaaagagga actgatcttc cggtactgca
     4981 gcggcagctg cgacgccgcc gagacaacct acgacaagat cctgaagaac ctgagccgga
     5041 accggcggct ggtgtccgac aaagtgggcc aggcctgctg cagacctatc gccttcgacg
     5101 acgacctgag cttcctggac gacaacctgg tgtaccacat cctgcggaag cacagcgcca
     5161 agagatgcgg ctgcatctga tgagctagct ctagacctgc agggtcgact ccggatgatc
     5221 agggccctgt acagatatcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5281 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5341 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5401 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5461 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5521 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5581 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5641 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5701 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5761 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5821 cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta caaggcagct
     5881 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5941 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6001 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6061 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6121 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6181 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6361 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6481 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     6541 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     6601 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6661 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6721 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6781 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6841 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6901 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6961 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     7021 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     7081 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     7141 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     7201 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7261 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7321 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7381 cacccagaaa cgctg
//
