LOCUS       sequence-263137-        8531 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of sfGFP tagged LifeAct probe to monitor actin 
            cytoskeleton.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8531)
  AUTHORS   Vjestica A, Marek M, Nkosi PJ, Merlini L, Liu G, Berard M, 
            Billault-Chaumartin I, Martin SG
  TITLE     A toolbox of stable integration vectors in the fission yeast 
            Schizosaccharomyces pombe.
  JOURNAL   J Cell Sci. 2020 Jan 8;133(1). pii: jcs.240754. doi: 
            10.1242/jcs.240754.
  PUBMED    31801797
REFERENCE   2  (bases 1 to 8531)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0782
FEATURES             Location/Qualifiers
     source          1..8531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2697..3152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3278..3300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3292..3309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3293..3309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3319..3338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3319..3337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      5013..5200
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5068..5092)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5275..5618
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6017..6214
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6271..6291)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6271..6289)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6310..6326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6310..6326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6323..6345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6334..6350
                     /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(6358..6364,6365..6382,6383..6388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6403..6424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6541..6558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6712..7300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6792..6811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7471..8262,8263..8331))
                     /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     8094..8113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8332..8436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8504..8522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaccg ggagaatcct gaataatgtg ctgtgaagca gttgaaagaa aattaaagaa
      241 cccaatataa tatgctataa agcaatcaaa aaataatagt aagaaaccca aataaaaccc
      301 cgttgaccat gttcccaatt aaattaacgc atattaatgc aaaaaatgca atattaagac
      361 tagttattaa accgataatt agtcttcaat tattaaaaga tgtagttgat ttgtattatc
      421 tttcaattgc taattctaaa tatattggag aattcaatga gtttatgcat tgaagtttaa
      481 gataacattg gcgctgattg cagcaaaagt aatcacgcat atagctgtag gaccaaaacg
      541 atcatcatat cggaccgcat tatgtttaaa cgttgcttta tatatggata acgacatata
      601 catttatttc ttgaaaaaat agcaaagcta tattgatttt agcgtatgct gctttaaagt
      661 tgcatttcac aatgcttgga aatgtaacga tgacagaatt aatacgatgc aaaactcaaa
      721 taataaactg cgcactaact cactacaata aacaactttg aaaaaaagat tcgttttttc
      781 aacatttacc atctcattaa gctgagctgc caaggtatat acatacttca tcgaatatga
      841 gcgaaaaaca ggttgtaggg atccttggag gtggtcaatt gggccgaatg atggtagagg
      901 cagcccatcg cttaaacatc aaatgcatca tcttggatgc agcaaattct cctgccaaac
      961 aaattgatgg aggacgtgag cacattgatg catcatttac tgaccccgat gcaattgttg
     1021 aactgtctaa gaagtgcacg ttattaacaa ctgaaattga gcatattaac actgatgcct
     1081 tggcagccgt tacgaaatct gttgctgttg aaccctctcc tgcaactctg cgatgcattc
     1141 aagacaaata tcttcaaaaa cagcatttac aggtttttaa gattgcactt cctgaatttt
     1201 gcgatgcacc tgaccaggaa agtgttgaaa aagcaggcca agagtttggt tatccttttg
     1261 tactgaaaag taaaacattg gcttacgacg gtcgtggaaa ttacgttgtt catcaaccat
     1321 ctgagattcc tactgccatc aaagcacttg gtgatcgtcc gctttatgtt gaaaagttcg
     1381 ttcctttctc catggaaatt gcagtgatgg tagtacgcag tttagacgga aaagtttatg
     1441 cttatcctac aactgagacc attcaaaagg ataatgtttg tcatttagta tatgcccctg
     1501 ctcgtcttcc cttctcaatt caacagcgtg ctcaaaccct tgccatggat gcggtgcgca
     1561 cttttgaagg cgctggtata tatggtgtag agatgtttgt tttgcaagat ggtgagacca
     1621 ttttactcaa cgaaattgct cctcggcctc acaattcagg tcactacacc attgaagctt
     1681 gcccaacttc tcagtttgaa gctcacttac gggccatatg tggtcttcct ttcagcgaaa
     1741 tcaacaccca actctcgact tccacaactc atgcgttgat ggtaaatatt ttaggtactg
     1801 atgatcctga ttatgtttca aagatcgcta aacgttctct gtccattccc ggtgcaactt
     1861 tgcatcttta tggtaaagct gaatctagaa agggtcgcaa gatgggacac gttaccatca
     1921 tttctgattc acctcaagaa tgtgaacgta ggtatcagat gcttcttgac gtcaaagatc
     1981 ctgtcgaatc acctgttgtt ggtattatca tgggttcgga ttctgattta agcaagatga
     2041 aagatgctgc cgtcatttta gatgaattca aggtgcctta cgaacttact attgtttcag
     2101 ctcaccgcac accagatcgc atggttactt atgctcgtac cgcagcttca agagggttgc
     2161 gtgtgattat tgctggtgct ggtggtgccg ctcatttgcc tggtatggtt gctgcaatga
     2221 cacctcttcc agtaatcggc gttcctgtaa aaggaagcac tcttgacgga gttgactctc
     2281 ttcactctat tgttcagatg cctcgaggtg tccctgtcgc cactgttgct atcaataata
     2341 gccaaaatgc cggtatttta gcctgtcgta tacttgctac atttcaaccc tcccttttgg
     2401 ctgctatgga gagctttatg gacaatatga aagagattgt tttagaaaag gctgataaat
     2461 tagaaaaagt tggttggaaa aattattctg cataggcgac catagacata actgttaaat
     2521 gattttgctt gctttttgtt ttaaataata ttttattttg aaattctaga ttgtaaaatg
     2581 tattctaaaa agtgattaaa aattaaaaat aaaaaccccc cgaataatgt gctgcgacgc
     2641 agctagggcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3181 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3301 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3361 ctcgaggtcg acgatctacg ataatgagac ggtgtttggg ttgtttttac tgaatttttc
     3421 ttttatttta tgacatctta atttttatgt taatgattat tcttatactt ttatctatct
     3481 ttgattaaaa ctaatattca ttcttttgat cactaattat ttttatagta tacgattcaa
     3541 cccaactata atacattgta tcaagagaaa tttttaatta gatgtaattt aatagccatt
     3601 tgataaaaaa aaattacctt tttttttgct agtgttaaat tattgatcat aacaatgtcg
     3661 aatgaatact aataaaatag ggacacgcga gttgcaaata cttgaaatac ctttgaagat
     3721 ttgtctgtga attttgataa gatttttatg accatctctt ttatattcga tatcgtgaac
     3781 tttcattttc ttttgttttt tgcttttcaa gttaaataac gcggaaatta tatttcctta
     3841 tatgtttata agagctttta aaggatttgc tatatcgctt acagtactac aaccggttgc
     3901 tcaatgttat ccgtttccgc ggtcttcttc cgtgcgcatt ctgccgtgaa gtgctaacgc
     3961 tgtgtgtggc tttatcaaaa aagtggctca aatatagcaa tttcatacat atttaaacac
     4021 ccattatact taaatgtata ttttaaaata tataagagat tttcactgta taactgtgta
     4081 tcttatctat atgtaatcaa cggcttcata ccacctcagc cagccgtgtt ataacttacc
     4141 gtttaccaac tacatttttt gtaacgaacc aaaaaaccct caaaagacaa gacccccggg
     4201 atgggtgtcg ctgaccttat caagaagttc gagtctattt ctaaggaagg atccggtggt
     4261 cccggtggtt taattaactc caagggtgaa gagctattta ctggggttgt acccattttg
     4321 gtagaactgg acggagatgt aaacggacat aaattctctg ttagaggtga gggcgaaggc
     4381 gatgccacca atggtaaatt gactctgaag tttatatgca ctacgggtaa attacctgtt
     4441 ccttggccaa ccctagtaac aactttgaca tatggtgttc aatgtttctc aagataccca
     4501 gaccatatga aaaggcatga tttctttaaa agtgctatgc cagaaggcta cgtgcaagag
     4561 agaactatct cctttaagga tgacggtacg tataaaacac gagcagaagt gaaattcgaa
     4621 ggggatacac tagttaatcg catcgaatta aagggtatag actttaagga agatggtaat
     4681 attctcggcc ataaacttga gtataatttc aactcgcata atgtgtacat tacagctgac
     4741 aaacaaaaga acggaattaa agcgaatttt aaaatcaggc acaacgtcga agatgggtct
     4801 gttcaacttg ccgatcatta tcagcaaaac acccctattg gtgatggtcc agtcttgtta
     4861 cccgataatc actacttaag cacacagtct agattgtcaa aagatccgaa tgaaaagcgt
     4921 gatcacatgg ttttattgga atttgtcacc gctgcaggaa taactcacgg aatggacgag
     4981 ctttataagt aaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     5041 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     5101 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     5161 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     5221 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     5281 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     5341 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     5401 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     5461 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     5521 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     5581 ggatacagtt ctcacatcac atccgaacat aaacaaccat gcctttgtct caagaagaat
     5641 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     5701 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     5761 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     5821 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     5881 gcggacggtg tcgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     5941 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     6001 gggagggcta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     6061 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     6121 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     6181 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     6241 tccagtttaa acgagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     6301 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8461 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8521 gccctttcgt c
//
