LOCUS       sequence-261161-        5271 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 5271)
  AUTHORS   Siegel PM, Shu W, Cardiff RD, Muller WJ, Massague J
  TITLE     Transforming growth factor beta signaling impairs Neu-induced 
            mammary tumorigenesis while promoting pulmonary metastasis.
  JOURNAL   Proc Natl Acad Sci U S A. 2003 Jul 8. 100(14):8430-5.
  PUBMED    12808151
REFERENCE   2  (bases 1 to 5271)
  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: "Proc Natl 
            Acad Sci U S A. 2003 Jul 8. 100(14):8430-5."
COMMENT     Sequence Label: pCMV5 HA-TBRII (delta Cyt)
FEATURES             Location/Qualifiers
     source          1..5271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1077..1103
                     /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    1633..2255
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1832..1851)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2279..2299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2444..2578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2630..2649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2631..2649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2663..2679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2663..2679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2676..2698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2687..2703
                     /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(2711..2717,2718..2735,2736..2741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2756..2777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2894..2911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3065..3653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3145..3164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3824..4615,4616..4684))
                     /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     4447..4466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4685..4789)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4816..5244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4903..4922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5113..5134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attcagatct ggtaccagct gctagcaagc ttgctagcgg
      961 ccgctcgagt cggtctatga cgagcagcgg ggtctgccat gggtcggggg ctgctcaggg
     1021 gcctgtggcc gctgcacatc gtcctgtgga cgcgtatcgc cagcacgatc ccaccgtatg
     1081 atgttcctga ttatgctagc ctccacgttc agaagtcggt taataacgac atgatagtca
     1141 ctgacaacaa cggtgcagtc aagtttccac aactgtgtaa attttgtgat gtgagatttt
     1201 ccacctgtga caaccagaaa tcctgcatga gcaactgcag catcacctcc atctgtgaga
     1261 agccacagga agtctgtgtg gctgtatgga gaaagaatga cgagaacata acgctagaga
     1321 cagtttgcca tgaccccaag ctcccctacc atgactttat tctggaagat gctgcttctc
     1381 caaagtgcat tatgaaggaa aaaaaaaagc ctggtgagac tttcttcatg tgttcctgta
     1441 gctctgatga gtgcaatgac aacatcatct tctcagaaga atataacacc agcaatcctg
     1501 acttgttgct agtcatattt caagtgacag gcatcagcct cctgccacca ctgggagttg
     1561 ccatatctgt catcatcatc ttctactgct accgcgttaa ccggcagcag aagctgagtt
     1621 catgaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     1681 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     1741 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     1801 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     1861 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     1921 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     1981 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     2041 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     2101 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     2161 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     2221 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg
     2281 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2341 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2401 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2581 ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga
     2641 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2701 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2761 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4801 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4861 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4921 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4981 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5041 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5101 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5161 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5221 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
