LOCUS       sequence-257657-        5739 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 5739)
  AUTHORS   Morales FC, Takahashi Y, Momin S, Adams H, Chen X, Georgescu MM
  TITLE     NHERF1/EBP50 head-to-tail intramolecular interaction masks 
            association with PDZ domain ligands.
  JOURNAL   Mol Cell Biol. 2007 Apr . 27(7):2527-37.
  PUBMED    17242191
REFERENCE   2  (bases 1 to 5739)
  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: "Mol Cell 
            Biol. 2007 Apr . 27(7):2527-37."
COMMENT     Sequence Label: pCMV-NHERF1-FL
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             660..683
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1805..2427
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2004..2023)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2451..2471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2456..2785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2636..2771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2823..2842)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2824..2842)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2856..2872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2856..2872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2869..2891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2880..2896
                     /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(2904..2910,2911..2928,2929..2934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2949..2970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3087..3104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3258..3846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3338..3357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4017..4808,4809..4877))
                     /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     4640..4659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4878..4982)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5009..5464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5096..5115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5306..5327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5590..5612
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5604..5621
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5605..5621
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaattga tctaccatgg
      661 actacaaaga cgatgacgac aagcttgcgg ccgcgatgag cgcggacgca gcggccgggg
      721 cgcccctgcc ccggctctgc tgcctggaga agggtccgaa cggctacggc ttccacctgc
      781 acggggagaa gggcaagttg ggccagtaca tccggctggt ggagcccggc tcgccggccg
      841 agaaggcggg gctgctggcg ggggaccggc tggtggaggt gaacggcgaa aacgtggaga
      901 aggagaccca ccagcaggtg gtgagccgca tccgcgccgc actcaacgcc gtgcgcctgc
      961 tggtggtcga ccccgagacg gacgagcagc tgcagaagct cggcgtccag gtccgagagg
     1021 agctgctgcg cgcccaggaa gcgccggggc aggccgagcc gccggccgcc gccgaggtgc
     1081 agggggctgg caacgaaaat gagcctcgcg aggccgacaa gagccacccg gagcagcgcg
     1141 agcttcggcc tcggctctgt accatgaaga agggccccag tggctatggc ttcaacctgc
     1201 acagcgacaa gtccaagcca ggccagttca tccggtcagt ggacccagac tccccggctg
     1261 aggcttcagg gctccgggcc caggatcgca ttgtggaggt gaacggggtc tgcatggagg
     1321 ggaagcagca tggggacgtg gtgtccgcca tcagggctgg cggggacgag accaagctgc
     1381 tggtggtgga cagggaaact gacgagttct tcaagaaatg cagagtgatc ccatctcagg
     1441 agcacctgaa tggtcccctg cctgtgccct tcaccaatgg ggagatacag aaggagagca
     1501 gtcgtgaagc cctggcagag gcagccttgg agagccccag gccagccctg gtgagatccg
     1561 cctccagtga caccagcgag gagctgaatt cccaagacag ccccccaaaa caggactcca
     1621 cagcgccctc gtctacctcc tcctccgacc ccatcctaga cttcaacatc tccctggcca
     1681 tggccaaaga gagggcccac cagaaacgca gcagcaaacg ggccccgcag atggactgga
     1741 gcaagaaaaa cgaactcttc agcaacctct gagttatcgg taccagtcga ctctagagga
     1801 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     1861 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     1921 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     1981 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     2041 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     2101 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     2161 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     2221 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     2281 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg
     2341 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg
     2401 gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg
     2461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2701 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2761 ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa
     2821 ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     2881 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2941 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3001 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3061 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3121 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3181 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3241 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3421 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3661 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3781 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3841 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3901 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3961 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4021 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4081 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4141 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4201 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4261 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4321 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4381 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4441 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4501 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4561 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4621 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4681 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4741 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4801 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4861 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4921 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4981 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     5041 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5101 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5161 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5221 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5281 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5341 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5401 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5461 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5521 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     5581 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat
     5641 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     5701 gctattggcc attgcatacg ttgtatctat atcataata
//
