LOCUS       sequence-140290-        6985 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 6985)
  AUTHORS   Liu C, Xu P, Lamouille S, Xu J, Derynck R
  TITLE     TACE-mediated ectodomain shedding of the type I TGF-beta receptor 
            downregulates TGF-beta signaling.
  JOURNAL   Mol Cell. 2009 Jul 10. 35(1):26-36.
  PUBMED    19595713
REFERENCE   2  (bases 1 to 6985)
  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: "Mol Cell. 
            2009 Jul 10. 35(1):26-36."
COMMENT     Sequence Label: pRK5M-ADAM10
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3192..3221
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3233..3242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3246..3380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3283..3302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3337..3356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3449..3806
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3449..3640
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3472..3492)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3657..3792
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3826..3843)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3826..3842)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3835..3857)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4055..4510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4142..4161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4352..4373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4527..4546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4646..4668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4706..4724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4792..4896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4897..4965,4966..5757)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSCGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5115..5134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5928..6516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6417..6436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6670..6687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6856..6877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6892..6897,6898..6915,6916..6922)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6930..6946
                     /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)."
     primer_bind     6935..6957
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6954..6970
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6954..6970
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attccccggg gatctcggaa gatggtgttg ctgagagtgt
      961 taattctgct cctctcctgg gcggcgggga tgggaggtca gtatgggaat cctttaaata
     1021 aatatatcag acattatgaa ggattatctt acaatgtgga ttcattacac caaaaacacc
     1081 agcgtgccaa aagagcagtc tcacatgaag accaattttt acgtctagat ttccatgccc
     1141 atggaagaca tttcaaccta cgaatgaaga gggacacttc ccttttcagt gatgaattta
     1201 aagtagaaac atcaaataaa gtacttgatt atgatacctc tcatatttac actggacata
     1261 tttatggtga agaaggaagt tttagccatg ggtctgttat tgatggaaga tttgaaggat
     1321 tcatccagac tcgtggtggc acattttatg ttgagccagc agagagatat attaaagacc
     1381 gaactctgcc atttcactct gtcatttatc atgaagatga tattaactat ccccataaat
     1441 acggtcctca ggggggctgt gcagatcatt cagtatttga aagaatgagg aaataccaga
     1501 tgactggtgt agaggaagta acacagatac ctcaagaaga acatgctgct aatggtccag
     1561 aacttctgag aaaaaaacgt acaacttcag ctgaaaaaaa tacttgtcag ctttatattc
     1621 agactgatca tttgttcttt aaatattacg gaacacgaga agctgtgatt gcccagatat
     1681 ccagtcatgt taaagcgatt gatacaattt accagaccac agacttctcc ggaatccgta
     1741 acatcagttt catggtgaaa cgcataagaa tcaatacaac tgctgatgag aaggacccta
     1801 caaatccttt ccgtttccca aatattggtg tggagaagtt tctggaattg aattctgagc
     1861 agaatcatga tgactactgt ttggcctatg tcttcacaga ccgagatttt gatgatggcg
     1921 tacttggtct ggcttgggtt ggagcacctt caggaagctc tggaggaata tgtgaaaaaa
     1981 gtaaactcta ttcagatggt aagaagaagt ccttaaacac tggaattatt actgttcaga
     2041 actatgggtc tcatgtacct cccaaagtct ctcacattac ttttgctcac gaagttggac
     2101 ataactttgg atccccacat gattctggaa cagagtgcac accaggagaa tctaagaatt
     2161 tgggtcaaaa agaaaatggc aattacatca tgtatgcaag agcaacatct ggggacaaac
     2221 ttaacaacaa taaattctca ctctgtagta ttagaaatat aagccaagtt cttgagaaga
     2281 agagaaacaa ctgttttgtt gaatctggcc aacctatttg tggaaatgga atggtagaac
     2341 aaggtgaaga atgtgattgt ggctatagtg accagtgtaa agatgaatgc tgcttcgatg
     2401 caaatcaacc agagggaaga aaatgcaaac tgaaacctgg gaaacagtgc agtccaagtc
     2461 aaggtccttg ttgtacagca cagtgtgcat tcaagtcaaa gtctgagaag tgtcgggatg
     2521 attcagactg tgcaagggaa ggaatatgta atggcttcac agctctctgc ccagcatctg
     2581 accctaaacc aaacttcaca gactgtaata ggcatacaca agtgtgcatt aatgggcaat
     2641 gtgcaggttc tatctgtgag aaatatggct tagaggagtg tacgtgtgcc agttctgatg
     2701 gcaaagatga taaagaatta tgccatgtat gctgtatgaa gaaaatggac ccatcaactt
     2761 gtgccagtac agggtctgtg cagtggagta ggcacttcag tggtcgaacc atcaccctgc
     2821 aacctggatc cccttgcaac gattttagag gttactgtga tgttttcatg cggtgcagat
     2881 tagtagatgc tgatggtcct ctagctaggc ttaaaaaagc aatttttagt ccagagctct
     2941 atgaaaacat tgctgaatgg attgtggctc attggtgggc agtattactt atgggaattg
     3001 ctctgatcat gctaatggct ggatttatta agatatgcag tgttcatact ccaagtagta
     3061 atccaaagtt gcctcctcct aaaccacttc caggcacttt aaagaggagg agacctccac
     3121 agcccattca gcaaccccag cgtcagcggc cccgagagag ttatcaaatg ggacacatga
     3181 gacgcgtcga cgagcagaag ctgatcagcg aggaggacct gctgtaagct tggccgccat
     3241 ggcccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3301 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3361 tatcttatca tgtctggatc gggaattaat tcggcgcagc accatggcct gaaataacct
     3421 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     3481 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3541 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3601 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3721 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3781 tttttggagg cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt
     3841 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3901 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3961 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4021 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4081 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4141 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4201 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4261 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4321 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4381 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttctgcctat
     4441 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4501 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4561 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4621 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4681 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4741 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4801 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5101 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5281 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5581 attgctgata aatctggagc cggtgagcgt gggtcttgcg gtatcattgc agcactgggg
     5641 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5701 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5761 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5821 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5881 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5941 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6001 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6061 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6121 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6181 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6241 ggctgaacgg ggggttcgtg cacacagccc agctttgagc gaacgaccta caccgaactg
     6301 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6361 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6421 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6481 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6541 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6601 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6661 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6721 ctccccgcgc gttggccgat tcattaatga gcgcccaata cgcaaaccgc ctctccccgc
     6781 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6841 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6901 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6961 cagctatgac atgattacga attaa
//
