LOCUS       sequence-80986-e        6316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiving vector for PUM-HD assembly and expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6316)
  AUTHORS   Abil Z, Denard CA, Zhao H
  TITLE     Modular assembly of designer PUF proteins for specific 
            post-transcriptional regulation of endogenous RNA.
  JOURNAL   J Biol Eng. 2014 Mar 1;8(1):7.
  PUBMED    24581042
REFERENCE   2  (bases 1 to 6316)
  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 Biol 
            Eng."; date: "2014-03-1"; volume: "8(1)"; pages: "7"
COMMENT     Sequence Label: pCMV-TTP(C147R)-GG-PUF
FEATURES             Location/Qualifiers
     source          1..6316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    20..642
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(219..238)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(664..684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        669..999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      850..985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1037..1056)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1038..1056)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1071..1087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1095..1111
                     /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(1119..1125,1126..1143,1144..1149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1164..1185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1302..1319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1472..2060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1552..1571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2231..3022,3023..3091))
                     /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     2854..2873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3092..3196)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3223..3651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3310..3329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3520..3541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3832..3848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4009..4388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4390..4589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4543..4563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4595..4660
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(5744..6070)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRFKQSTLDLIKDPARPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGVSSGN
                     LPQL"
     primer_bind     5953..5969
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6066..6082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6090..6106
                     /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(6114..6120,6121..6138,6139..6144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6159..6180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtcgactcta gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
       61 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
      121 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
      181 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
      241 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
      301 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
      361 gtttttttgg tagagacggg gtttcaccat attggccagg ctgaactcca actcctaatc
      421 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
      481 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
      541 gcataggcta cctgccatgg cccaaccggt gggacatttg agttgcttgc ttggcactgt
      601 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttc
      661 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
      721 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaaagtcccc aggctcccca
      781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      901 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa
     1021 aaccagaaag ttaattccct atagtgagtc gtattaaatt cgtaatcatg gtcatagctg
     1081 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1141 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1201 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1441 aggaaccgta aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     1681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     1921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2341 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
     2401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     3241 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3301 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3361 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     3421 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3481 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3541 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3601 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3661 tattaacaaa atattaacgt ttacaatttc cccattcgcc attcaggctg cgcaactgtt
     3721 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3781 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3841 cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc atattagtca
     3901 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     3961 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat
     4021 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4081 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4141 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4201 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4261 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4321 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4381 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
     4441 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4501 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
     4561 gtgtacggtg ggaggtctat ataagcccac catggactac aaagaccatg acggtgatta
     4621 taaagatcat gacatcgatt acaaggatga cgatgacaag atggatctga ctgccatcta
     4681 cgagagcctc ctgtcgctga gccctgacgt gcccgtgcca tccgaccatg gagggactga
     4741 gtccagccca ggctggggct cctcgggacc ctggagcctg agcccctccg actccagccc
     4801 gtctggggtc acctcccgcc tgcctggccg ctccaccagc ctagtggagg gccgcagctg
     4861 tggctgggtg cccccacccc ctggcttcgc accgctggct ccccgcctgg gccctgagct
     4921 gtcaccctca cccacttcgc ccactgcaac ctccaccacc ccctcgcgct acaagactga
     4981 gctatgtcgg accttctcag agagtgggcg ctgccgctac ggggccaagt gccagtttgc
     5041 ccatggcctg ggcgagctgc gccaggccaa tcgccacccc aaatacaaga cggaactccg
     5101 tcacaagttc tacctccagg gccgctgccc ctacggctct cgctgccact tcatccacaa
     5161 ccctagcgaa gacctggcgg ccccgggcca ccctcctgtg cttcgccaga gcatcagctt
     5221 ctccggcctg ccctctggcc gccggacctc accaccacca ccaggcctgg ccggcccttc
     5281 cctgtcctcc agctccttct cgccctccag ctccccacca ccacctgggg accttccact
     5341 gtcaccctct gccttctctg ctgcccctgg cacccccctg gctcgaagag atcccacccc
     5401 agtctgttgc ccctcctgcc gaagggccac tcctatcagc gtctgggggc ccttgggtgg
     5461 cctggttcgg accccctctg tacagtccct gggatccgac cctgatgaat atgccagcag
     5521 cggcagcagc ctggggggct ctgactctcc cgtcttcgag gcgggagttt ttgcaccacc
     5581 ccagcccgtg gcagcccccc ggcgactccc catcttcaat cgcatctctg tttctgaggg
     5641 aggtggtgga agcggaggtg gcggttcagg tggcggtgga tctggaggcg gtgggggccg
     5701 cagccgcctt ttggaagatt ttcgaaacaa ccggtacccc aatttacaac tgcgggagat
     5761 tgccggaaga gaccccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     5821 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5881 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     5941 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg gggcgcgccg
     6001 gatccttaat taagtctaga gtcgactgtt taaacctgca ggcatgcaag cttggcgtaa
     6061 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6121 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6181 attgcgttgc gctcactgcc cgctttccag tcggtctcaa ccccacatcg caactcttcg
     6241 taagtacacc tatggcaagc acattctggc caagctggag aagtactaca tgaagaacgg
     6301 tgttgactta gggtaa
//
