LOCUS       sequence-199722-        7274 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 7274)
  AUTHORS   Zhang J, Vinuela A, Neely MH, Hallett PJ, Grant SG, Miller GM, 
            Isacson O, Caron MG, Yao WD
  TITLE     Inhibition of the dopamine D1 receptor signaling by PSD-95.
  JOURNAL   J Biol Chem. 2007 May 25. 282(21):15778-89.
  PUBMED    17369255
REFERENCE   2  (bases 1 to 7274)
  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 
            Chem. 2007 May 25. 282(21):15778-89."
COMMENT     Sequence Label: PSD-95 FLAG
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        490..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     644..664
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    695..713
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    716..734
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     744..768
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      865..874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3049..3072
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3101..3118)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3107..3331
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3464..3483)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3615..3662
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3670..4692
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3670..4692
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4822..4943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4859..4878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4913..4932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4992..5008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4992..5008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5005..5027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5016..5032
                     /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(5040..5046,5047..5064,5065..5070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5085..5106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5223..5240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5394..5982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5474..5493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6153..6944,6945..7013))
                     /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     6776..6795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7014..7118)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7193..7212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct
      721 atcagtgata gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      781 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
      841 gcgtttaaac ttaagcttgg taccgccacc atggactgtc tctgtatagt gacaaccaag
      901 aaataccgct accaagatga agacacgccc cctctggaac acagcccggc ccacctcccc
      961 aaccaggcca attctccccc tgtgattgtc aacacggaca ccctagaagc cccaggatat
     1021 gagttgcagg tgaatggaac agagggggag atggagtatg aggagatcac attggaaagg
     1081 ggtaactcag gtctgggctt cagcatcgca ggtggcgctg acaacccgca catcggtgac
     1141 gacccgtcca tttttatcac caagatcatt cctggtgggg ctgcagccca ggatggccgc
     1201 ctcagggtca atgacagcat cctgtttgta aatgaagtgg atgttcggga ggtgacccat
     1261 tcagctgcgg tggaggccct caaagaggca ggttccatcg ttcgcctcta tgtcatgcgc
     1321 cggaaacccc cagccgaaaa ggtcatggag atcaaactca tcaaagggcc taaaggactt
     1381 ggcttcagca ttgcgggggg cgttgggaac cagcacatcc ctggagataa cagcatctat
     1441 gtaacgaaga tcatcgaagg aggtgctgcc cacaaggatg gcaggttgca gattggagac
     1501 aagatcctgg cggtcaacag tgtggggctg gaggacgtca tgcacgagga tgccgtggca
     1561 gccctgaaga acacatatga cgttgtgtac ctaaaggtgg ccaagcccag caatgcctac
     1621 ctgagtgaca gctatgctcc cccagacatc acaacctcgt attctcagca cctggacaat
     1681 gagatcagtc atagcagcta cttgggcact gactacccca cagccatgac ccccacttcc
     1741 cctcggcgct actcccctgt ggccaaggac ctgctggggg aggaagacat tccccgggaa
     1801 ccaaggcgga tcgtgatcca tcggggctcc accggcctgg gcttcaacat cgtgggcggc
     1861 gaggatggtg aaggcatctt catctccttc atccttgctg ggggtccagc cgacctcagt
     1921 ggggagctac ggaaggggga ccagatcctg tcggtcaatg gtgttgacct ccgcaatgcc
     1981 agtcacgaac aggctgccat tgccctgaag aatgcgggtc agacggtcac gatcatcgct
     2041 cagtataaac cagaagagta tagtcgattc gaggccaaga tccatgatct tcgggaacag
     2101 ctcatgaata gtagcctagg ctcagggact gcatccttgc gaagcaaccc caagaggggc
     2161 ttctacatta gggccctgtt tgattacgac aagaccaagg actgcggttt cttgagccag
     2221 gccctgagct tccgcttcgg ggatgtgctt catgtcattg acgctggtga cgaagagtgg
     2281 tggcaagcac ggcgggtcca ctccgacagt gagaccgacg acattggctt cattcccagc
     2341 aaacggcggg tcgagcgacg agagtggtca aggttaaagg ccaaggactg gggctccagc
     2401 tctggatcac agggtcgaga agactcggtt ctgagctatg agacggtgac ccagatggaa
     2461 gtgcactatg ctcgtcccat catcatcctt ggacccacca aagaccgtgc caacgatgat
     2521 cttctctccg agttccccga caagtttgga tcctgtgtcc ctcatacgac acgtcctaag
     2581 cgggaatatg agatagacgg ccgggattac cactttgtct cctcccggga gaaaatggag
     2641 aaggacatcc aggcacacaa gttcattgag gctggccagt acaacagcca cctctatggg
     2701 accagcgtcc agtctgtgcg agaggtagca gagcagggga agcactgcat cctcgatgtc
     2761 tcggccaatg ccgtgcggcg gctgcaggcg gcccacctgc accccatcgc catcttcatc
     2821 cgtccccgct ccctggagaa tgtgctagag atcaataagc ggatcacaga ggagcaagcc
     2881 cggaaagcct tcgacagagc cacgaagctg gagcaggagt tcacagagtg cttctcagcc
     2941 atcgtagagg gcgacagctt tgaagagatc tatcacaaag tgaaacgtgt cattgaagac
     3001 ctctcaggcc cctacatctg ggtcccagcc cgagagagac tcctcgagga ctacaaggac
     3061 gacgatgaca agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3601 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     3661 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     3721 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     3781 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     3841 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     3901 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     3961 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     4021 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     4081 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     4141 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     4201 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     4261 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     4321 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     4381 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     4441 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     4501 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     4561 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     4621 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     4681 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4861 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4921 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5161 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5221 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5281 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5941 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6001 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6061 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6241 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6301 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6361 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6421 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6481 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6541 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6601 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6661 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6721 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6781 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6841 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6901 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6961 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7021 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7081 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7141 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7201 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7261 agtagtgcgc gagc
//
