LOCUS       sequence-151504-        6265 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 6265)
  TITLE     Joan Massague lab plasmids
REFERENCE   2  (bases 1 to 6265)
  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-HA with various GS domain mutations
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        235..253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(292..621)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     606..626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    650..1272
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1054..1073
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(1283..1309)
                     /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"
     CDS             complement(1289..1315)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2977..3180)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3006..3026)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3181..3560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3721..3738)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3721..3737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3730..3752)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3878..4333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4015..4036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4227..4246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4360..4464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4465..4533,4534..5325)
                     /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     complement(4683..4702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5496..6084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5985..6004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6238..6255
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
      241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
      301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
      361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
      661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
      721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
      781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
      841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
      901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
      961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     1261 gggatgccac ccgggatccc tagaggctag cataatcagg aacatcatac ggatacattt
     1321 tgatgccttc ctgttgactg agttgcgata atgttttctt aatccgcaat gctgtaagcc
     1381 tagctgctcc attggcatac caacattctc tcataatttt agccattact ctcaaggctt
     1441 cacagctctg ccatctgttt gggatatttg gccttaactt ctgttcacaa acaacttttc
     1501 tcatttcttc aactgatggg tcagaaggta caagatcata ataaggcagt tggtaatctt
     1561 catgaattcc accaatggaa catcgtcgag caatttccca gaatactaag cccattgcat
     1621 agatgtcagc acgtttgaag gattcaaaat gtttcatatt tatggaatca tcgagaactt
     1681 caggggccat gtaccttttt gttcccactc tgtggtttgg agcaatatca atggtatctg
     1741 tggctgaatc atgtcttact gccagtccta agtctgcaat acagcaagtt ccattcttct
     1801 ttaccaagat attctttgat ttcaaatctc tatgagcaat ggctggcttt ccttgggtac
     1861 caacaatctc catgtgaaga tgggcaagac cgctcgccgt ggacagagca agttttatca
     1921 ttccttccac agtaactgtg tatctgttta agtaatcaaa aagagatcca tgctcatgat
     1981 aatctgacac caaccagagc tgagtccaag taccattgtc tttattgtct gctgctataa
     2041 atcccaggat gttttcatga cgtaacatta cagtttgata aatctctgcc tcacggaacc
     2101 acgaacgttc ttctctagag gagaatatct taacagcaac ttcttctccc cgccactttc
     2161 ctctccaaac ttctccaaat cgacctttgc caatgctttc ttgtaacaca atatctctcg
     2221 caattgttct ctgaacaagc aatggtaaac ctgagccaga acctgacgtt gtcatatcat
     2281 aaattaagtc tttcaacgta gtaccctctg aaataaaagg gcgatctaat gaagggtcct
     2341 cttcatttgg cactcgatgg tgaatgacag tgcggttgtg gcagatatag accatcaaca
     2401 tgagtgagat gcagacgaag cacactggtc cagcaatgac agctgccagt tccacaggac
     2461 caaggccagg tgatgacttt acagtagttg gaagttctat tttattgcaa tggtcctgat
     2521 tgcagcaata tgttgtagtc acagacccag tttttgaaga gggtgcacat acaaacggcc
     2581 tatctcgagg aattaagtca atttcagcta tacacatgct gttgtgtata actttgtctg
     2641 tggtctctgt gacagagaca aagcagagcc catctgtcac acaagtaaaa ttgtcttttg
     2701 tacagaggtg gcagaaacac tgtaacgccg tcgcccccgg gagcagcgcc gccgccgccg
     2761 ccgccgccgc cgccagcacg aggaggagca gccggggacg cggagcagcg accgccgcct
     2821 ccatggtccc gccgccaccg cctgtggccc ggcccggccc ggccgcgccg ctgcctcacc
     2881 ccagcaaacc tcgcctcgcc cgacagcaac ggaattcgat atcaagctta tcgatacgcg
     2941 tggtaccaga tctgaattct gacggttcac taaacgagct ctgcttatat agacctccca
     3001 ccgtacacgc ctaccgccca tttgcgtcaa cggggcgggg ttattacgac attttggaaa
     3061 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     3121 aaatccccgt gagtcaaacc gctatccacg cccattggtg tactgccaaa accgcatcac
     3181 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccgtaaggt
     3241 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgg
     3301 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3361 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3421 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3481 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3541 actagtcaat aatcaatgtc aacatggcgg tcatattgga catgagccaa tataaatgta
     3601 catattatga tatagataca acgtatgcaa tggccaatag ccaatattga tttatgctat
     3661 ataaccaatg actaatatgg ctaattgcca atattgattc aatgtataga tcagcttggc
     3721 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3781 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3841 cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg
     3901 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3961 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4021 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4081 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4141 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4201 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4261 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4321 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4381 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4441 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4501 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4561 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4621 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4681 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4741 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4801 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4861 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4921 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4981 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5041 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5101 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5161 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5221 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5281 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5341 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5401 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5461 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5521 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5581 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5641 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5701 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5761 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5821 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5881 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5941 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6001 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6061 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6121 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6181 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6241 gagtcagtga gcgaggaagc ggaag
//
