LOCUS       sequence-449148-        5301 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 5301)
  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 5301)
  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-TIMP3
FEATURES             Location/Qualifiers
     source          1..5301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        226..605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     760..780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     806..830
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1040..1058
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1040..1057
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1071..1090
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1772..1801
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1813..1822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1826..1960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1863..1882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1917..1936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2029..2386
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2029..2220
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2052..2072)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2237..2372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2406..2423)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2406..2422)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2415..2437)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2635..3090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2722..2741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2932..2953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3107..3126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3226..3248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3286..3304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3372..3476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3477..3545,3546..4337)
                     /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     complement(3695..3714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4508..5096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4997..5016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5250..5267
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgacatg attacgaatt aattcgagct cgcccgacat tgattattga
      241 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      781 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      841 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
      901 gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt accgcctata
      961 gagtctatag gcccaccccc ttggcttcgt tagaacgcgg ctacaattaa tacataacct
     1021 tatgtatcat acacatacga tttaggtgac actatagaat aacatccact ttgcctttct
     1081 ctccacaggt gtccactccc aggtccaact gcacctcggt tctatcgatg caatgacccc
     1141 ttggctcggg ctcatcgtgc tcctgggcag ctggagcctg ggggactggg gcgccgaggc
     1201 gtgcacatgc tcgcccagcc acccccagga cgccttctgc aactccgaca tcgtgatccg
     1261 ggccaaggtg gtggggaaga agctggtaaa ggaggggccc ttcggcacgc tggtctacac
     1321 catcaagcag atgaagatgt accgaggctt caccaagatg ccccatgtgc agtacatcca
     1381 cacggaagct tccgagagtc tctgtggcct taagctggag gtcaacaagt accagtacct
     1441 gctgacaggt cgcgtctatg atggcaagat gtacacgggg ctgtgcaact tcgtggagag
     1501 gtgggaccag ctcaccctct cccagcgcaa ggggctgaac tatcggtatc acctgggttg
     1561 taactgcaag atcaagtcct gctactacct gccttgcttt gtgacttcca agaacgagtg
     1621 tctctggacc gacatgctct ccaatttcgg ttaccctggc taccagtcca aacactacgc
     1681 ctgcatccgg cagaagggcg gctactgcag ctggtaccga ggatgggccc ccccggataa
     1741 aagcatcatc aatgccacag accccgtcga cgagcagaag ctgatcagcg aggaggacct
     1801 gctgtaagct tggccgccat ggcccaactt gtttattgca gcttataatg gttacaaata
     1861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1921 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat tcggcgcagc
     1981 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     2041 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2341 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc
     2401 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2461 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2521 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     2581 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2761 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2821 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2881 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2941 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3001 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3061 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3121 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3301 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3661 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3781 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4021 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4081 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4321 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4621 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5221 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5281 gcccaatacg caaaccgcct c
//
