LOCUS       sequence-352644-        6106 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 6106)
  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 6106)
  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-Smad5
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..84
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     76..93
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     77..93
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        254..633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        634..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     788..808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2383..3005
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2582..2601)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3029..3049)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3034..3363
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3214..3349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3401..3420)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3402..3420)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3434..3450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3434..3450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3447..3469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3458..3474
                     /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(3482..3488,3489..3506,3507..3512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3527..3548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3665..3682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3916..3935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4595..5386,5387..5455))
                     /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     5218..5237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5456..5560)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5587..6042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5674..5693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5884..5905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
       61 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac attgaatcaa
      121 tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg
      181 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
      241 tgaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      301 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      361 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      421 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      481 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
      541 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
      601 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat
      661 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      721 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
      781 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      841 ttagtgaacc gtcagaattc agatctggta ccacgcgtat cgataagctt gcatgcctgc
      901 aggtcgactc tagaggatcc gcgatgacgt caatggccag cttgttttct ttcactagtc
      961 cagccgtgaa gcgattgttg ggctggaaac aaggtgacga ggaagagaaa tgggcagaaa
     1021 aggcagtgga tgctttagtg aaaaagctga agaagaagaa gggtgctatg gaggagctgg
     1081 agaaagcctt gagcagccca ggacagccaa gcaagtgtgt cacgatcccc aggtccttgg
     1141 atggacgtct gcaagtttct cacaggaaag gcttgcccca tgttatatat tgccgtgttt
     1201 ggcgctggcc agatttgcag agccatcacg agctaaaacc attggatatt tgtgaatttc
     1261 cttttggatc taagcaaaag gaagtttgta tcaatccata ccactataag agagtggaga
     1321 gtccagtctt acctccagta ttagtgcctc gtcacaatga attcaatcca caacacagcc
     1381 ttctggttca gttcaggaac ctgagccaca atgaaccgca catgccacaa aacgccacgt
     1441 ttcccgattc tttccaccaa cccaacaacg ctcctttccc cttatctcct aacagcccct
     1501 atcctccttc ccctgctagc agcacatatc ccaactcccc agcaagctct ggacctggaa
     1561 gtccatttca actcccagct gacacccctc cccctgccta tatgccacct gatgatcaga
     1621 tggccccaga taattcccag cctatggata caagcagtaa catgattcct cagaccatgc
     1681 ccagcatatc cagcagagat gttcagcctg tcgcctatga ggagcccaaa cactggtgtt
     1741 cgattgtcta ctatgaatta aacaatcgtg ttggggaagc ttttcatgca tcttctacta
     1801 gtgtgttagt agatggattt acagatcctt caaataacaa aagtagattc tgcctgggat
     1861 tgttgtcaaa tgttaatcgt aattcaacta ttgaaaacac taggcggcat attggaaaag
     1921 gtgttcatct atactacgtt ggtggggagg tgtacgctga gtgtcttagt gacagcagca
     1981 tctttgttca gagtaggaac tgcaactttc accatggctt ccatcccacc accgtctgta
     2041 agatccccag cagctgcagc ctcaagattt ttaacaatca ggagtttgct cagcttctgg
     2101 ctcagtcagt caaccatgga ttcgaggctg tgtatgagct caccaagatg tgtaccattc
     2161 gaatgagctt tgtcaagggc tggggagcag agtaccaccg acaggacgtc accagtactc
     2221 cctgctggat tgagattcac ctccacgggc ctctgcagtg gctggataaa gtccttactc
     2281 agatgggctc tccgctgaac cccatttctt ctgtttcata gtgcagaagt attctttcaa
     2341 ctatattttt agtggacttg ttttaatttt agaggggatc ccgggtggca tccctgtgac
     2401 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2461 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2521 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2581 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2641 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2701 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2761 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2821 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac
     2881 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat
     2941 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt
     3001 gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3181 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3241 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3301 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3361 aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta
     3421 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5581 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5641 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5701 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5761 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5821 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5881 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5941 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6001 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
     6061 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctatta
//
