LOCUS       sequence-315030-        6300 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 6300)
  AUTHORS   Macias-Silva M, Hoodless PA, Tang SJ, Buchwald M, Wrana JL
  TITLE     Specific activation of Smad1 signaling pathways by the BMP7 type I 
            receptor, ALK2.
  JOURNAL   J Biol Chem. 1998 Oct 2. 273(40):25628-36.
  PUBMED    9748228
REFERENCE   2  (bases 1 to 6300)
  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: "J Biol 
            Chem. 1998 Oct 2. 273(40):25628-36."
COMMENT     Sequence Label: pCMV5-ActRIIA
FEATURES             Location/Qualifiers
     source          1..6300
                     /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             2287..2313
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    2324..2946
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2523..2542)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2970..2990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2975..3304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3155..3290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3342..3361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3343..3361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3375..3391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3375..3391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3388..3410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3399..3415
                     /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(3423..3429,3430..3447,3448..3453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3468..3489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3606..3623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3777..4365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3857..3876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4536..5327,5328..5396))
                     /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     5159..5178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5397..5501)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5528..5983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5615..5634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5825..5846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6109..6131
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6123..6140
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6124..6140
                     /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 agaattcaga tctggtacca cgcgtatcga taagcttgca tgcctgcagg tcgagcggcc
      661 gccagtgtgc tctaaagctc cgaggaagac ccagggaact ggatatctag cgagaacttc
      721 ctacggcttc tccggcgcct cgggaaaatg ggagctgctg caaagttggc gttcgccgtc
      781 tttcttatct cttgctcttc aggtgctata cttggcagat cagaaactca ggagtgtctt
      841 ttctttaatg ctaattggga aagagacaga accaaccaga ctggtgttga accttgctat
      901 ggtgataaag ataaacggcg acattgtttt gctacctgga agaatatttc tggttccatt
      961 gaaatagtga agcaaggttg ttggctggat gatatcaact gctatgacag gactgattgt
     1021 atagaaaaaa aagacagccc tgaagtgtac ttttgttgct gtgagggcaa tatgtgtaat
     1081 gaaaagttct cttattttcc ggagatggaa gtcacacagc ccacttcaaa tcctgttaca
     1141 ccgaagccac cctattacaa cattctgctg tattccttgg taccactaat gttaattgca
     1201 ggaattgtca tttgtgcatt ttgggtgtac agacatcaca agatggccta ccctcctgta
     1261 cttgttccta ctcaagaccc aggaccaccc ccaccttccc cattactagg gttgaagcca
     1321 ttgcagctgt tagaagtgaa agcaagggga agatttggtt gtgtctggaa agcccagttg
     1381 ctcaatgaat atgtggctgt caaaatattt ccaatacagg acaaacagtc ctggcagaat
     1441 gaatatgaag tctatagtct acctggaatg aagcatgaga acatactaca gttcattggt
     1501 gcagagaaaa gaggcaccag tgtggatgtg gacctgtggc taatcacagc atttcatgaa
     1561 aagggctcac tgtcagactt tcttaaggct aatgtggtct cttggaatga actttgtcat
     1621 attgcagaaa ccatggctag aggattggca tatttacatg aggatatacc tggcttaaaa
     1681 gatggccaca agcctgcaat ctctcacagg gacatcaaaa gtaaaaatgt gctgttgaaa
     1741 aacaatctga cagcttgcat tgctgacttt gggttggcct taaagttcga ggctggcaag
     1801 tctgcaggtg acacccatgg gcaggttggt acccggaggt atatggctcc agaggtgttg
     1861 gagggtgcta taaacttcca aagggacgca tttctgagga tagatatgta cgccatggga
     1921 ttagtcctat gggaattggc ttctcgttgc actgctgcag atggacccgt agatgagtac
     1981 atgttaccat ttgaggaaga aattggccag catccatctc ttgaagatat gcaggaagtt
     2041 gttgtgcata aaaaaaagag gcctgtttta agagattatt ggcagaaaca tgcaggaatg
     2101 gcaatgctct gtgaaacgat agaagaatgt tgggatcatg atgcagaagc caggttatca
     2161 gctggatgtg taggtgaaag aattactcag atgcaaagac taacaaatat cattactaca
     2221 gaggacattg taacagtggt cacaatggtg acaaatgttg actttcctcc caaagaatct
     2281 agtctatatg atgttcctga ttatgctagc ctctagggat cccgggtggc atccctgtga
     2341 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     2401 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     2461 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     2521 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     2581 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     2641 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     2701 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     2761 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata
     2821 ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca
     2881 tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt
     2941 tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3241 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3301 caaaaagctc ctcgaggaac tgaaaaacca gaaagttaat tccctatagt gagtcgtatt
     3361 aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5521 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5581 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5641 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5701 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5761 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5821 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5881 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5941 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc
     6001 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6061 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6121 gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag
     6181 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     6241 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
