LOCUS       Exported                6256 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV5 TBRI-His
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 Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6256
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     59..81
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     73..90
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     74..90
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     785..805
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2496..2513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2530..3152
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2729..2748)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3176..3196)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3181..3510
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3361..3496
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3423..3442
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3548..3567)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3549..3567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3581..3597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3581..3597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3594..3616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3605..3621
                     /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(3629..3659)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3674..3695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3812..3829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3983..4571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4063..4082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4742..5602)
                     /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     5365..5384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5603..5707)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5734..6189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5821..5840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6031..6052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
       61 cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat
      121 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
      181 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
      241 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      841 gtgaaccgtc agaattcaga tctggtacca cgcgtatcga taagcttgat atcgaattcc
      901 gttgctgtcg ggcgaggcga ggtttgctgg ggtgaggcag cggcgcggcc gggccgggcc
      961 gggccacagg cggtggcggc gggaccatgg aggcggcggt cgctgctccg cgtccccggc
     1021 tgctcctcct cgtgctggcg gcggcggcgg cggcggcggc ggcgctgctc ccgggggcga
     1081 cggcgttaca gtgtttctgc cacctctgta caaaagacaa ttttacttgt gtgacagatg
     1141 ggctctgctt tgtctctgtc acagagacca cagacaaagt tatacacaac agcatgtgta
     1201 tagctgaaat tgacttaatt cctcgagata ggccgtttgt atgtgcaccc tcttcaaaaa
     1261 ctgggtctgt gactacaaca tattgctgca atcaggacca ttgcaataaa atagaacttc
     1321 caactactgt aaagtcatca cctggccttg gtcctgtgga actggcagct gtcattgctg
     1381 gaccagtgtg cttcgtctgc atctcactca tgttgatggt ctatatctgc cacaaccgca
     1441 ctgtcattca ccatcgagtg ccaaatgaag aggacccttc attagatcgc ccttttattt
     1501 cagagggtac tacgttgaaa gacttaattt atgatatgac aacgtcaggt tctggctcag
     1561 gtttaccatt gcttgttcag agaacaattg cgagaactat tgtgttacaa gaaagcattg
     1621 gcaaaggtcg atttggagaa gtttggagag gaaagtggcg gggagaagaa gttgctgtta
     1681 agatattctc ctctagagaa gaacgttcgt ggttccgtga ggcagagatt tatcaaactg
     1741 taatgttacg tcatgaaaac atcctgggat ttatagcagc agacaataaa gacaatggta
     1801 cttggactca gctctggttg gtgtcagatt atcatgagca tggatctctt tttgattact
     1861 taaacagata cacagttact gtggaaggaa tgataaaact tgctctgtcc acggcgagcg
     1921 gtcttgccca tcttcacatg gagattgttg gtacccaagg aaagccagcc attgctcata
     1981 gagatttgaa atcaaagaat atcttggtaa agaagaatgg aacttgctgt attgcagact
     2041 taggactggc agtaagacat gattcagcca cagataccat tgatattgct ccaaaccaca
     2101 gagtgggaac aaaaaggtac atggcccctg aagttctcga tgattccata aatatgaaac
     2161 attttgaatc cttcaaacgt gctgacatct atgcaatggg cttagtattc tgggaaattg
     2221 ctcgacgatg ttccattggt ggaattcatg aagattacca actgccttat tatgatcttg
     2281 taccttctga cccatcagtt gaagaaatga gaaaagttgt ttgtgaacag aagttaaggc
     2341 caaatatccc aaacagatgg cagagctgtg aagccttgag agtaatggct aaaattatga
     2401 gagaatgttg gtatgccaat ggagcagcta ggcttacagc attgcggatt aagaaaacat
     2461 tatcgcaact cagtcaacag gaaggcatca aaatgcatca ccaccatcat cactaaacta
     2521 gtggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2581 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2641 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2701 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2761 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2821 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2881 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     2941 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3001 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     3061 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     3121 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
     3181 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3241 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3301 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3361 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3421 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3481 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa
     3541 gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt
     3601 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3661 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3721 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3781 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3841 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3901 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3961 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4021 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4081 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4141 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4201 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4261 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4321 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4381 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4441 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4501 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4561 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4621 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4681 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4741 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4801 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4861 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4921 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4981 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5041 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5101 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5161 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5221 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5281 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5341 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5401 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5461 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5521 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5581 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5641 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5701 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     5761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     6181 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6241 ctcttcgcta ttacgc
//
