LOCUS       sequence-139066-        8621 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 8621)
  AUTHORS   Tsukazaki T, Chiang TA, Davison AF, Attisano L, Wrana JL
  TITLE     SARA, a FYVE domain protein that recruits Smad2 to the TGFbeta 
            receptor.
  JOURNAL   Cell. 1998 Dec 11. 95(6):779-91.
  PUBMED    9865696
REFERENCE   2  (bases 1 to 8621)
  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: "Cell. 1998 
            Dec 11. 95(6):779-91."
COMMENT     Sequence Label: pCMV5B-FLAG-SARA
FEATURES             Location/Qualifiers
     source          1..8621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2580..2597
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2581..2597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2758..3137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3138..3341
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3292..3312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3397..3420
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    7402..8024
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7601..7620)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(8048..8068)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8053..8382
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8233..8368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8420..8439)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8421..8439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8453..8469)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8453..8469)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8466..8488)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8477..8493
                     /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(8501..8507,8508..8525,8526..8531))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8546..8567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     3361 attgatctgg tacgtaccac gcgtatcgat accatggact acaaggacga cgatgacaag
     3421 ggtcgactgt ggattgatga aaatgctgtt gcagaagacc agttaattaa gagaaactat
     3481 agttgggatg atcaatgcag tgctgttgaa gtgggagaga agaaatgtgg aaacctggct
     3541 tgtctgccag atgagaagaa tgttcttgtt gtagccgtca tgcataactg tgataaaagg
     3601 acattacaaa acgatttaca ggattgtaat aattataata gtcaatccct tatggatgct
     3661 tttagctgtt cactggataa tgaaaacaga caaactgatc aatttagttt tagtataaat
     3721 gagtccactg aaaaagatat gaattcagag aaacaaatgg atccattgaa tagaccgaaa
     3781 acagagggga gatctgttaa ccatctgtgt cctacttcat ctgatagtct agccagtgtc
     3841 tgttcccctt cacaattaaa ggatgacgga agtataggta gagacccctc catgtctgcg
     3901 attacaagtt taacggttga ttcagtaatc tcatcccagg gaacagatgg atgtcctgct
     3961 gttaaaaagc aagagaacta tataccagat gaggacctca ctggcaaaat cagctctcct
     4021 aggacagatc tagggagtcc aaattccttt tcccacatga gtgaggggat tttgatgaaa
     4081 aaagagccag cagaggagag caccactgaa gaatccctcc ggtctggttt acctttgctt
     4141 ctcaaaccag acatgcctaa tgggtctgga aggaataatg actgtgaacg gtgttcagat
     4201 tgccttgtgc ctaatgaagt tagggctgat gaaaatgaag gttatgaaca tgaagaaact
     4261 cttggcacta cagaattcct taatatgaca gagcatttct ctgaatctca ggacatgact
     4321 aattggaagt tgactaaact aaatgagatg aatgatagcc aagtaaacga agaaaaggaa
     4381 aagtttctac agattagtca gcctgaggac actaatggtg atagtggagg acagtgtgtt
     4441 ggattggcag atgcaggtct agatttaaaa ggaacttgca ttagtgaaag tgaagaatgt
     4501 gatttctcca ctgttataga cacaccagca gcaaattatc tatctaatgg ttgtgattcc
     4561 tatggaatgc aagacccagg tgtttctttt gttccaaaga ctttaccctc caaagaagat
     4621 tcagtaacag aagaaaaaga aatagaggaa agcaagtcag aatgctactc aaatatttat
     4681 gaacagagag gaaatgaggc cacagaaggg agtggactac ttttaaacag cactggtgac
     4741 ctaatgaaga aaaattattt acataatttc tgtagtcaag ttccatcagt gcttgggcaa
     4801 tcttccccca aggtagtagc aagcctgcca tctatcagtg ttccttttgg tggtgcaaga
     4861 cccaagcaac cttctaatct taaacttcaa attccaaagc cattatcaga ccatttacaa
     4921 aatgactttc ctgcaaacag tggaaataat actaaaaata aaaatgatat tcttgggaaa
     4981 gcaaaattag gggaaaactc agcaaccaat gtatgcagtc catctttggg aaacatctct
     5041 aatgtcgata caaatgggga acatttagaa agttatgagg ctgagatctc cactagacca
     5101 tgccttgcat tagctccaga tagcccagat aatgatctca gagctggtca gtttggaatt
     5161 tctgccagaa agccattcac cactctgggt gaggtggctc cagtatgggt accggattct
     5221 caggctccaa attgcatgaa atgtgaagcc aggtttacat tcaccaaaag gaggcatcac
     5281 tgcagagcat gtgggaaggt tttctgtgct tcctgctgta gcctgaaatg taaactgtta
     5341 tacatggaca gaaaggaagc tagagtgtgt gtaatctgcc attcagtgct aatgaatgct
     5401 caagcctggg agaacatgat gagtgcctca agccagagcc ctaaccctaa caatcctgct
     5461 gaatactgtt ctactatccc tcccttgcag caagctcagg cctcaggagc tctgagctct
     5521 ccacctccca ctgtgatggt acctgtggga gttttaaagc accctggagc agaagtggct
     5581 cagcccagag agcagaggcg agtttggttt gctgatggga tcttgcccaa tggagaagtt
     5641 gctgatgcag ccaaattaac aatgaatgga acttcctctg caggaaccct ggctgtgtca
     5701 cacgacccag tcaagccagt aactaccagt cctctaccag cagagacgga tatttgtcta
     5761 ttctctggga gtataactca ggttggaagt cctgttggaa gtgcaatgaa tcttattcct
     5821 gaagatggcc ttcctcccat tctcatctcc actggtgtaa aaggagacta tgctgtggaa
     5881 gagaaaccat cacagatttc agtaatgcag cagttggagg atggtggccc tgacccactt
     5941 gtatttgttt taaatgcaaa tttgttgtca atggttaaaa ttgtaaatta tgtgaacagg
     6001 aagtgctggt gtttcacaac caagggaatg catgcagtgg gtcagtctga gatagtcatt
     6061 cttctacagt gtttaccgga tgaaaagtgt ttgccaaagg atatctttaa tcactttgtg
     6121 cagctttatc gggatgctct ggcagggaat gtggtgagca acttgggaca ttccttcttc
     6181 agtcaaagtt tccttggcag taaagaacat ggtggattct tatatgtgac atctacctac
     6241 cagtcactgc aagacctagt actcccaacc ccaccttact tgtttgggat tcttatccag
     6301 aaatgggaaa ctccttgggc taaagtattt cctatccgtc tgatgttgag acttggagct
     6361 gaatatcgac tttatccatg cccactattc agtgtcagat ttcggaagcc attgtttgga
     6421 gagacggggc ataccatcat gaatcttctt gcagacttca gaaattacca gtataccttg
     6481 ccagtagttc aaggtttggt ggttgatatg gaagttcgga aaactagcat caaaattccc
     6541 agcaacagat acaatgagat gatgaaagcc atgaacaagt ccaatgagca tgtcctggca
     6601 ggaggtgcct gcttcaatga aaaggcagac tctcatcttg tgtgtgtaca gaatgatgat
     6661 ggaaactatc agacccaggc tatcagtatt cacaatcagc ccagaaaagt gactggtgcc
     6721 agtttctttg tgttcagtgg cgctctgaaa tcctcttctg gataccttgc caagtccagt
     6781 attgtggaag atggtgttat ggtccagatt actgcagaga acatggattc cttgaggcag
     6841 gcactgcgag agatgaagga cttcaccatc acctgtggga aggcggacgc ggaggaaccc
     6901 caggagcaca tccacatcca gtgggtggat gatgacaaga acgttagcaa gggtgtcgta
     6961 agtcctatag atgggaagtc catggagact ataacaaatg tgaagatatt ccatggatca
     7021 gaatataaag caaatggaaa agtaatcaga tggacagagg tgtttttcct agaaaacgat
     7081 gaccagcaca attgcctcag tgatcctgca gatcacagta gattgactga gcatgttgcc
     7141 aaagcttttt gccttgctct ctgtcctcac ctgaaacttc tgaaggaaga tggaatgacc
     7201 aaactgggac tacgtgtgac acttgactca gatcaggttg gctatcaagc agggagcaat
     7261 ggccagcccc ttccctcgca gtacatgaat gatctggaca gcgccttggt gccggtgatc
     7321 catggagggg cctgccagct tagtgagggc cccgttgtca tggaactcat cttttatatt
     7381 ctggaaaaca tcgtataacc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     7441 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     7501 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     7561 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     7621 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     7681 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     7741 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     7801 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     7861 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     7921 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     7981 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct
     8041 tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     8101 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     8161 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     8221 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     8281 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     8341 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg
     8401 aaaaaccaga aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc
     8461 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     8521 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     8581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct g
//
