LOCUS       sequence-137799-        5891 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 5891)
  AUTHORS   Varelas X, Sakuma R, Samavarchi-Tehrani P, Peerani R, Rao BM, 
            Dembowy J, Yaffe MB, Zandstra PW, Wrana JL
  TITLE     TAZ controls Smad nucleocytoplasmic shuttling and regulates human 
            embryonic stem-cell self-renewal.
  JOURNAL   Nat Cell Biol. 2008 Jul . 10(7):837-48.
  PUBMED    18568018
REFERENCE   2  (bases 1 to 5891)
  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: "Nat Cell 
            Biol. 2008 Jul . 10(7):837-48."
COMMENT     Sequence Label: 3XFlag pCMV5-TOPO TAZ (S89A)
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             647..712
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1937..2559
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2136..2155)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2583..2603)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2588..2917
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2768..2903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2959..2978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2960..2978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2992..3008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2992..3008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3005..3027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3016..3032
                     /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(3040..3046,3047..3064,3065..3070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3223..3240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3394..3982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3474..3493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4153..4944,4945..5013))
                     /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     4776..4795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5014..5118)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5145..5600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5232..5251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5442..5463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5726..5748
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5740..5757
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5741..5757
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct cgtttagtga accgtcagaa ttgatctggt accatggact acaaagacca
      661 tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca agcttgggaa
      721 tccggcctcg gcgccccctc cgctcccgcc gcctgggcag caagtgatcc acgtcacgca
      781 ggacctagac acagacctcg aagccctctt caactctgtc atgaatccga agcctagctc
      841 gtggcggaag aagatcctgc cggagtcttt ctttaaggag cctgattcgg gctcgcactc
      901 gcgccagtcc agcaccgact cgtcgggcgg ccacccgggg cctcgactgg ctgggggtgc
      961 ccagcatgtc cgctcgcacg cctcgccagc gtccctgcag ctgggcaccg gcgcgggtgc
     1021 tgcgggtagc cccgcgcagc agcacgcgca cctccgccag cagtcctacg acgtgaccga
     1081 cgagctgcca ctgcccccgg gctgggagat gaccttcacg gccactggcc agaggtactt
     1141 cctcaatcac atagaaaaaa tcaccacatg gcaagaccct aggaaggcga tgaatcagcc
     1201 tctgaatcat atgaacctcc accctgccgt cagttccaca ccagtgcctc agaggtccat
     1261 ggcagtatcc cagccaaatc tcgtgatgaa tcaccaacac cagcagcaga tggcccccag
     1321 taccctgagc cagcagaacc accccactca gaacccaccc gcagggctca tgagtatgcc
     1381 caatgcgctg accactcagc agcagcagca gcagaaactg cggcttcaga gaatccagat
     1441 ggagagagaa aggattcgaa tgcgccaaga ggagctcatg aggcaggaag ctgccctctg
     1501 tcgacagctc cccatggaag ctgagactct tgccccagtt caggctgctg tcaacccacc
     1561 cacgatgacc ccagacatga gatccatcac taataatagc tcagatcctt tcctcaatgg
     1621 agggccatat cattcgaggg agcagagcac tgacagtggc ctggggttag ggtgctacag
     1681 tgtccccaca actccggagg acttcctcag caatgtggat gagatggata caggagaaaa
     1741 cgcaggacaa acacccatga acatcaatcc ccaacagacc cgtttccctg atttccttga
     1801 ctgtcttcca ggaacaaacg ttgacttagg aactttggaa tctgaagacc tgatccccct
     1861 cttcaatgat gtagagtctg ctctgaacaa aagtgagccc tttctaacct ggctgtaaga
     1921 attcaagggc gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     1981 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     2041 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     2101 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     2161 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     2221 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     2281 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     2341 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     2401 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     2461 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     2521 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     2581 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2641 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2761 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2821 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2881 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgatc gaggaactga
     2941 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     3001 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3061 aaagtgtaaa gcctggggtg cctaatgagc gagctaactc acattaattg cgttgcgctc
     3121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4141 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4201 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4261 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4321 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4381 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4441 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4501 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4561 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4621 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4681 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4741 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4801 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4861 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4921 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4981 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5041 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5101 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     5161 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5221 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5281 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5341 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5401 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5461 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5521 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5581 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5641 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5701 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     5761 aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     5821 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     5881 atttatattg g
//
