LOCUS       sequence-38674-e        7073 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 7073)
  AUTHORS   Weinman EJ, Wang Y, Wang F, Greer C, Steplock D, Shenolikar S
  TITLE     A C-terminal PDZ motif in NHE3 binds NHERF-1 and enhances cAMP 
            inhibition of sodium-hydrogen exchange.
  JOURNAL   Biochemistry. 2003 Nov 4. 42(43):12662-8.
  PUBMED    14580213
REFERENCE   2  (bases 1 to 7073)
  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: 
            "Biochemistry. 2003 Nov 4. 42(43):12662-8."
COMMENT     Sequence Label: pcDNA-N1-PDZ2-GAGA
FEATURES             Location/Qualifiers
     source          1..7073
                     /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        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             907..929
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             940..957
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             967..984
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             991..1035
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1051..1065
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(2498..2515)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2504..2728
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2774..3202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2861..2880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3071..3092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3211..3231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3216..3545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3396..3531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3458..3477
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3594..4619
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3594..4619
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4749..4870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4786..4805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4840..4859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4919..4935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4919..4935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4932..4954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4943..4959
                     /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(4967..4973,4974..4991,4992..4997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5012..5033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5150..5167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5321..5906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5401..5420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6077..6868,6869..6937))
                     /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     6700..6719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6938..7042)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctaga
      901 aataattttg tttaacttta agaaggagat atacatatgc accatcatca tcatcattct
      961 tctggtctgg tgccacgcgg ttctggtatg aaagaaaccg ctgctgctaa attcgaacgc
     1021 cagcacatgg acagcccaga tctgggtacc gacgacgacg acaaggccat ggctgatcgc
     1081 gccgctgagc cgtcgcgggg cgagatgagc gcggacgcgg cggccggggc gcccctgccc
     1141 cggctctgct gcctggagaa ggggccgaac ggctacggct tccacctgca tggggagaag
     1201 ggcaaggtgg gccagtacat ccggctggtg gagcccggct ctccggccga gaaggcgggg
     1261 ctgctggccg gggaccggct ggtggaggtg aacggcgaga acgtggagaa ggagacccac
     1321 cagcaggtgg tgagccgcat ccgcgccgcg ctcaacgccg tgcgcctgct ggtggtcgac
     1381 cccgacacgg acgagcagtt caggaagctc ggagtccaga tccgggggga gctgctacgc
     1441 gcccaggcgg ggccggagca ggccgggccg ccggccgccc ccggcgagca gggcccggcc
     1501 ggtgagaacg agccgcgcga ggtcgagaag agccacccgg agcggcgcga gctccggcct
     1561 cggctctgcg ccatgaagaa gggccacaac ggcgccggcg ccaacctgca cagcgacaag
     1621 tccaggccgg gccagttcat ccgcgccgtg gacccagact ccccggctga ggcctcgggg
     1681 ctccgggagc aggaccgcat cgtggaggtg aacggggtct gcgtggaggg caagcagcat
     1741 ggggacgtgg tgactgccat caaggccggc ggggatgagg ccaagctgct ggtggtggac
     1801 aaggagaccg acgagttctt caagaaatgc aaagtggtcc catcttctga gcacctcaac
     1861 ggtcccttgc ctgagccctt caccaatggg gagatacaga agaacaaccc tgagaccctg
     1921 gccccagcgg cctcggagag ccccaggcca gccctggcaa gatccgcctc cagtgacacc
     1981 agcgaggagc tggcttccca agacagcccc aagaaagagg actccacggc cccctcctcg
     2041 acctcctcct cctccgaccc catcctggac ttcagcatct cgctggccgt ggccaaggag
     2101 agggcccacc agaagcgcag cagcagacgg gccccccaaa tggactggag cgagaaaaag
     2161 gagcttttca gcaacctctg agcgccccgg ccgcccaccc cccgtcacct cctctccgcc
     2221 cgattctcca gtccccagcc gcccaccctc cttcacctcc tctccgccca cttctccaat
     2281 ccctggccgc ccaccccacc tcacctcctc tccgcccgct tctccagtcc ccggccgccc
     2341 accctacctc acctcctctc cgccccgctt ctccaatcac cctcgaatcg atctacagat
     2401 cagcaccagt gccccgttcc tggccatgtg acttttttag agaactgtgc acttccctgc
     2461 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2521 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2581 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2641 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2701 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2761 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3181 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3361 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3481 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3541 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgatgaaaa
     3601 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     3661 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     3721 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     3781 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     3841 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     3901 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     3961 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     4021 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     4081 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     4141 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     4201 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     4261 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     4321 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     4381 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     4441 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     4501 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     4561 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
     4621 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4681 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4741 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4801 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4861 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4921 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4981 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5041 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5101 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5761 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5821 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6001 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6661 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
