LOCUS       sequence-364368-        6256 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 6256)
  TITLE     Joan Massague lab plasmids
REFERENCE   2  (bases 1 to 6256)
  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"
COMMENT     Sequence Label: pCMV5 TBRI-His
FEATURES             Location/Qualifiers
     source          1..6256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             834..851
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    868..1490
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1067..1086)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1514..1534)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1519..1848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1699..1834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1886..1905)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1887..1905)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1919..1935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1919..1935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1932..1954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1943..1959
                     /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(1967..1973,1974..1991,1992..1997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2012..2033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2150..2167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2321..2909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2401..2420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3080..3871,3872..3940))
                     /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     3703..3722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3941..4045)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4072..4527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4159..4178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4369..4390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4653..4675
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4667..4684
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4668..4684
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4845..5224
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5225..5428
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5379..5399
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gagaagaagt tgctgttaag atattctcct ctagagaaga acgttcgtgg ttccgtgagg
       61 cagagattta tcaaactgta atgttacgtc atgaaaacat cctgggattt atagcagcag
      121 acaataaaga caatggtact tggactcagc tctggttggt gtcagattat catgagcatg
      181 gatctctttt tgattactta aacagataca cagttactgt ggaaggaatg ataaaacttg
      241 ctctgtccac ggcgagcggt cttgcccatc ttcacatgga gattgttggt acccaaggaa
      301 agccagccat tgctcataga gatttgaaat caaagaatat cttggtaaag aagaatggaa
      361 cttgctgtat tgcagactta ggactggcag taagacatga ttcagccaca gataccattg
      421 atattgctcc aaaccacaga gtgggaacaa aaaggtacat ggcccctgaa gttctcgatg
      481 attccataaa tatgaaacat tttgaatcct tcaaacgtgc tgacatctat gcaatgggct
      541 tagtattctg ggaaattgct cgacgatgtt ccattggtgg aattcatgaa gattaccaac
      601 tgccttatta tgatcttgta ccttctgacc catcagttga agaaatgaga aaagttgttt
      661 gtgaacagaa gttaaggcca aatatcccaa acagatggca gagctgtgaa gccttgagag
      721 taatggctaa aattatgaga gaatgttggt atgccaatgg agcagctagg cttacagcat
      781 tgcggattaa gaaaacatta tcgcaactca gtcaacagga aggcatcaaa atgcatcacc
      841 accatcatca ctaaactagt ggatcccggg tggcatccct gtgacccctc cccagtgcct
      901 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
      961 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     1021 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     1081 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     1141 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     1201 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     1261 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     1321 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     1381 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     1441 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     1501 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1561 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1621 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1681 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1741 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1801 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag
     1861 gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt
     1921 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1981 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2041 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2101 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2761 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2821 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4081 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4141 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4201 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4261 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4321 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4381 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4441 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4501 ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga
     4561 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     4621 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     4681 cagtgccaag ctgatctata cattgaatca atattggcaa ttagccatat tagtcattgg
     4741 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     4801 tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac
     4861 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     4921 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     4981 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5041 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     5101 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     5161 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     5221 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     5281 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     5341 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     5401 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagaatt cagatctggt
     5461 accacgcgta tcgataagct tgatatcgaa ttccgttgct gtcgggcgag gcgaggtttg
     5521 ctggggtgag gcagcggcgc ggccgggccg ggccgggcca caggcggtgg cggcgggacc
     5581 atggaggcgg cggtcgctgc tccgcgtccc cggctgctcc tcctcgtgct ggcggcggcg
     5641 gcggcggcgg cggcggcgct gctcccgggg gcgacggcgt tacagtgttt ctgccacctc
     5701 tgtacaaaag acaattttac ttgtgtgaca gatgggctct gctttgtctc tgtcacagag
     5761 accacagaca aagttataca caacagcatg tgtatagctg aaattgactt aattcctcga
     5821 gataggccgt ttgtatgtgc accctcttca aaaactgggt ctgtgactac aacatattgc
     5881 tgcaatcagg accattgcaa taaaatagaa cttccaacta ctgtaaagtc atcacctggc
     5941 cttggtcctg tggaactggc agctgtcatt gctggaccag tgtgcttcgt ctgcatctca
     6001 ctcatgttga tggtctatat ctgccacaac cgcactgtca ttcaccatcg agtgccaaat
     6061 gaagaggacc cttcattaga tcgccctttt atttcagagg gtactacgtt gaaagactta
     6121 atttatgata tgacaacgtc aggttctggc tcaggtttac cattgcttgt tcagagaaca
     6181 attgcgagaa ctattgtgtt acaagaaagc attggcaaag gtcgatttgg agaagtttgg
     6241 agaggaaagt ggcggg
//
