LOCUS       sequence-7280-ex        8497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8497)
  AUTHORS   Marr RA, Guan H, Rockenstein E, Kindy M, Gage FH, Verma I, Masliah 
            E, Hersh LB
  TITLE     Neprilysin regulates amyloid Beta peptide levels.
  JOURNAL   J Mol Neurosci. 2004  . 22(1-2):5-11.
  PUBMED    14742905
REFERENCE   2  (bases 1 to 8497)
  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: "J Mol 
            Neurosci. 2004  . 22(1-2):5-11."
COMMENT     Sequence Label: pCSC-SP-PW (aka: pBOB)
FEATURES             Location/Qualifiers
     source          1..8497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1056..1181
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1674..1907
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2092..2136
                     /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    2388..2505
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2528..2831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2832..3035
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3032..3056
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3164..3180)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3165..3181)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    3303..3891
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3356..3376)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3774..3785)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4298..4478
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4504..4521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4510..4734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4780..5208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5077..5098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5217..5237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5222..5552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5403..5538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5465..5484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5600..5647
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5666..6040
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6170..6291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6207..6226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6261..6280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6340..6356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6340..6356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6353..6375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6364..6380
                     /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(6388..6394,6395..6412,6413..6418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6433..6454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6571..6588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6742..7330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6822..6841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7501..8292,8293..8361))
                     /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     8124..8143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8362..8466)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cctgaaagcg aaagggaaac cagagctctc tcgacgcagg actcggcttg
     1081 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1141 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1201 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1261 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1321 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1381 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1441 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1501 agaccaccgc acagcaagcg gccgctgatc ttcagacttg gaggaggaga tatgagggac
     1561 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1621 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1681 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcctc aatgacgctg
     1741 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1801 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccaa
     1861 gcaagaatcc tagctgtgga aagataccta aaggatcaac agctcctagg gatttggggt
     1921 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1981 tctctggaac agatctggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     2041 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     2101 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2161 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2221 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2281 cagacccacc tcccaatccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2341 ggagagagag acagagacag atccattcga ttagtgaacg gatcaacttt taaaagaaaa
     2401 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2461 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga taagcttggg
     2521 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2581 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2641 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2701 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     2761 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2821 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     2881 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     2941 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     3001 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     3061 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctag aggatccacc
     3121 ggtggccgcc tgggcccgtt tgagaattcg atatcaagct tatcgatacc gtcgacctcg
     3181 agtttaaacc tggaaaaaca tggagcaatc acaagtagca acacagcagc taccaatgct
     3241 gcttgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     3301 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3361 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3421 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3481 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3541 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3601 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3661 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     3721 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3781 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3841 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc
     3901 gagctcggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     3961 aaaagaaaag gggggactgg aagggctaat tcactcccaa agaagacaag atatccttga
     4021 tctgtggatc taccacacac aaggctactt ccctgattga cagaactaca caccagggcc
     4081 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     4141 taagatagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     4201 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     4261 tcatcacgtg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     4321 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     4381 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     4441 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gccgctttaa acccgctgat
     4501 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4561 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4621 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4681 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4741 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4921 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4981 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5101 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     5161 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     5221 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     5281 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5341 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5401 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5461 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5521 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5581 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     5641 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     5701 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     5761 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     5821 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     5881 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     5941 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     6001 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     6061 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6301 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6901 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7141 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7621 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8461 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
