LOCUS       sequence-137800-        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 WT
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             621..686
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1911..2533
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2110..2129)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2557..2577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2562..2891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2742..2877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2933..2952)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2934..2952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2966..2982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2966..2982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2979..3001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2990..3006
                     /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(3014..3020,3021..3038,3039..3044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3197..3214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3368..3956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3448..3467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4127..4918,4919..4987))
                     /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     4750..4769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4988..5092)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5119..5574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5206..5225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5416..5437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5700..5722
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5714..5731
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5715..5731
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc tggtaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      661 tcgattacaa ggatgacgat gacaagcttg ggaatccggc ctcggcgccc cctccgctcc
      721 cgccgcctgg gcagcaagtg atccacgtca cgcaggacct agacacagac ctcgaagccc
      781 tcttcaactc tgtcatgaat ccgaagccta gctcgtggcg gaagaagatc ctgccggagt
      841 ctttctttaa ggagcctgat tcgggctcgc actcgcgcca gtccagcacc gactcgtcgg
      901 gcggccaccc ggggcctcga ctggctgggg gtgcccagca tgtccgctcg cactcgtcgc
      961 ccgcgtccct gcagctgggc accggcgcgg gtgctgcggg tagccccgcg cagcagcacg
     1021 cgcacctccg ccagcagtcc tacgacgtga ccgacgagct gccactgccc ccgggctggg
     1081 agatgacctt cacggccact ggccagaggt acttcctcaa tcacatagaa aaaatcacca
     1141 catggcaaga ccctaggaag gcgatgaatc agcctctgaa tcatatgaac ctccaccctg
     1201 ccgtcagttc cacaccagtg cctcagaggt ccatggcagt atcccagcca aatctcgtga
     1261 tgaatcacca acaccagcag cagatggccc ccagtaccct gagccagcag aaccacccca
     1321 ctcagaaccc acccgcaggg ctcatgagta tgcccaatgc gctgaccact cagcagcagc
     1381 agcagcagaa actgcggctt cagagaatcc agatggagag agaaaggatt cgaatgcgcc
     1441 aagaggagct catgaggcag gaagctgccc tctgtcgaca gctccccatg gaagctgaga
     1501 ctcttgcccc agttcaggct gctgtcaacc cacccacgat gaccccagac atgagatcca
     1561 tcactaataa tagctcagat cctttcctca atggagggcc atatcattcg agggagcaga
     1621 gcactgacag tggcctgggg ttagggtgct acagtgtccc cacaactccg gaggacttcc
     1681 tcagcaatgt ggatgagatg gatacaggag aaaacgcagg acaaacaccc atgaacatca
     1741 atccccaaca gacccgtttc cctgatttcc ttgactgtct tccaggaaca aacgttgact
     1801 taggaacttt ggaatctgaa gacctgatcc ccctcttcaa tgatgtagag tctgctctga
     1861 acaaaagtga gccctttcta acctggctgt aagaattcaa gggcgatccc gggtggcatc
     1921 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     1981 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2041 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2101 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2161 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2221 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2281 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2341 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     2401 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     2461 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     2521 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2821 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2881 gcttttgcaa aaagctcctc gatcgaggaa ctgaaaaacc agaaagttaa ttccctatag
     2941 tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     3001 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3061 gagcgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3121 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3181 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3241 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3301 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3361 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3421 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3481 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3541 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3601 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3661 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3721 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3781 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3841 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3901 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3961 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4021 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4081 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4141 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4201 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4261 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4321 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4381 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4441 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4501 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4561 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4621 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4681 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4741 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4801 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4861 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4921 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4981 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5041 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5101 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5161 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5221 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5281 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5341 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5401 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5461 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5521 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     5581 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5641 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5701 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat tgaatcaata
     5761 ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg gctattggcc
     5821 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
     5881 accgccatgt t
//
