LOCUS       sequence-249009-        6346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis          .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6346)
  AUTHORS   Zhu H, Kavsak P, Abdollah S, Wrana JL, Thomsen GH
  TITLE     A SMAD ubiquitin ligase targets the BMP pathway and affects 
            embryonic pattern formation.
  JOURNAL   Nature. 1999 Aug 12. 400(6745):687-93.
  PUBMED    10458166
REFERENCE   2  (bases 1 to 6346)
  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: "Nature. 
            1999 Aug 12. 400(6745):687-93."
COMMENT     Sequence Label: pCS2(+)-XSmurf1C699A
FEATURES             Location/Qualifiers
     source          1..6346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    2398..2532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2422..2441)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2476..2495
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2645..2665)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2645..2663)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2684..2700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2684..2700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2697..2719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2708..2724
                     /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(2732..2738,2739..2756,2757..2762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2915..2932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3086..3674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3166..3185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3845..4636,4637..4705))
                     /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     4468..4487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4706..4810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4836..5291)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4973..4994)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5185..5204
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5362..6346
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg cggccgcgtc gaccagcctt
      121 ccggcgtttg gagggatgtc gaacgttgtg actcggagag gtggatcgag tatcagagtt
      181 cgtctgacag ttttatgtgc aaagaacttg gcaaagaggg attttttcag attacctgat
      241 ccatttgcta aaatagttgt ggatggatct ggtcagtgcc actcaacaga cactgtaaag
      301 aacacattgg atccaaagtg gaatcagcat tatgacctgt atgttggtaa aatggattca
      361 atcacaatca gtatctggaa tcataagaaa attcacaaga aacagggagc tggcttccta
      421 ggctgtgtga gactattatc caatgctatt agcagattga aagacacagg ctaccagcgc
      481 ttggatctat gcaaattaaa ccccacagac aatgacgctg tgcggggcca aattgtagtc
      541 agtttacaga cacgagatag gatcggtaca ctaggttctg ttgttgattg cagaggactg
      601 ctagacaatg aaggggcact tttagaagat actggccctg gcaggcccct tagttgtttc
      661 atggatgagc cagcacctta tactgatgga ccaggtgcag ctggaggagg acccggccgg
      721 ctagtagaat ctccaggtca agaacagaga ctccaggcac agagagttcg tggaccagaa
      781 gtcagagagc atgtccagac acctcaaaat cgatcccatg ggttccagtc acaggacttg
      841 cctgaaggtt atgaacaaag aactacagtt caggggcagg tgtatttctt gcacacacaa
      901 acgggtgtca gcacatggca tgatccccga ataccaagag accttaacag cgtgaattgt
      961 gatgaccttg gctctctgcc cgctggatgg gaggtcagaa ccactgtatc tggaagaata
     1021 tactttgtag atcacaataa tcgaacaaca cagttcacag acccaagact acatcacatc
     1081 ataaatcatc agtcccagct aaaggagcca aatcatgcta taccagtgca gagtgatgga
     1141 tctttagaag atggagatga gtttcctgct cagagatatg agagagactt ggtacagaaa
     1201 cttaaagtcc tgaggcatga gttatcattg ctgcagcctc aagcagggca ctgccgtgtt
     1261 gaagtgtcaa gagaagaaat atttgaggag tcctatagac aaataatgaa gatgagaccc
     1321 aaagatctga agaaaaggtt gatggtgaaa tttcgtggag aagaaggact tgattatggt
     1381 ggagttgcca gggagtggct gtatttacta tgtcatgaaa tgctgaatcc atattatgga
     1441 ctcttccagt attctacaga taatatttat actctgcaaa ttaatcctga ttcatctatt
     1501 aatccagatc atttgtccta tttccatttt gttggtcgaa ttatgggatt ggctgtattt
     1561 catggacatt acattaatgg ggggttcaca gttcctttct acaagcaact tttggggaag
     1621 cccattcagt tgtctgacct ggaatctgtg gatccagagt tgcataaaag cctagtttgg
     1681 attttggaga atgacattac ttcagtatta gaccacacat tctgtgttga gcacaatgca
     1741 tttggccgac ttctccagca tgaattaaaa ccaaatggca agaacctgca agtaacagaa
     1801 gagaacaaga aagagtatgt gaggttgtat gttaattgga ggttcatgag aggtatagag
     1861 gctcagttct tagctctgca gaaaggtttt aatgagctga taccacaaca tctgttgaag
     1921 ccatttgagc aaaaagaact agagcttatt atcggaggct tggataaaat tgacatcagt
     1981 gactggaagg caaacacaag attaaaacac tgcttagcta atagcaatat tgttcagtgg
     2041 ttctggcaag cagtggagtc gtttgatgag gagagaaggg ccagactact tcagttcgtc
     2101 acaggctcta cacgtgttcc tcttcaaggc tttaaagctc ttcaaggttc tacaggtgct
     2161 gctggaccca gactttttac tattcatttg attgatgcta acacagataa cctcccaaag
     2221 gctcacacag cctttaaccg aattgacatt cctccttatg aatcgtatga aaagctctat
     2281 gagaaattat taacagctgt agaagagact agtggttttg ctgtggaata aatgtcgacg
     2341 cggccgcgaa ttcaaggcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat
     2401 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2461 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2521 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2581 tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     2641 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     2701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta
     2761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4801 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     4861 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4921 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4981 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5041 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5101 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5161 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5221 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5281 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5341 ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc
     5401 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     5461 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     5521 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
     5581 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     5641 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     5701 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     5761 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     5821 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     5881 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     5941 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     6001 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     6061 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     6121 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     6181 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     6241 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     6301 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
