LOCUS       sequence-263761-        8380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP targeted to the plasma membrane with an 
            amphipathic helix of RitC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8380)
  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 8380)
  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: "J Cell 
            Sci."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0605
FEATURES             Location/Qualifiers
     source          1..8380
                     /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"
     CDS             1640..2434
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      complement(2926..3381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3275..3294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3507..3529
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3521..3538
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3522..3538
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3548..3567
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3548..3566
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6120..6140)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6120..6138)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6159..6175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6159..6175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6172..6194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6183..6199
                     /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(6207..6213,6214..6231,6232..6237))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6252..6273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6390..6407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6561..7149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6641..6660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7320..8111,8112..8180))
                     /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     7943..7962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8181..8285)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8353..8371
                     /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 ggtattagtc aaaactagta tactttttct cggagtataa tacacaatat
      241 cggtgcaaat aggttttaaa attgctccaa tcacatgttc ttaagaaaaa aacgtcaaaa
      301 gaaatctaag tattcccaaa ataattcctc ctgacatgag aagaataaaa aacactaaga
      361 tgcattgtca gtatgaaaca caagccaaag ttgctaacaa taggatcaat tcttacttac
      421 tggcaatgtt acgaggaatc ttggtaggag gatgtttggc aacataacgt tcttttgaat
      481 ttgtttttac acgacgagta atgttcttct gaaactgtgt gtttgcacga cgctgagagg
      541 gagtttgaac cgtctaaaag acatagttag gatccatgat gcacagattt ctaaagataa
      601 aactacttac aggcatgttg gtttattaaa aaagaaaaac aggaatgata agagggaaag
      661 aatacgtgta cagagcgctt acataatgag tgagtgtcat tattgaggaa tgccatgtca
      721 gatttgacac aactcagaac ttattaactt tttatgattg gaacttttgt tttgcccaag
      781 gaatttttaa tggaagaagt tgttttataa atgatattga atttatatcc actcttttta
      841 gcgcatttgc atctctactt taggttcttc tttgactaga ttctaggatg cgtattggat
      901 atactggtct ttgaatcatt gtcgaaaaaa aaattcatgg ttgtacccat gagcaaactg
      961 cttttgtggt tatccatgtg cttcttttgt tcagcatcta tcttcttaat cgcatggaag
     1021 atttgattta aaccaaaaga aatattgtta aactttgaca gaagctttaa attttgattc
     1081 catacattaa atggataaaa ctcctgaagc ttagctacaa atcccactgg ctatatgtat
     1141 gcatttgtgt taaaaaagtt tgtatagatt atttaatcta ctcagcattc tttctctaaa
     1201 taggaatttg ttacttaatg gagaaaaaaa tgtttcgatt tacctagtgt atttgtttgt
     1261 atactcacgt ttaatttcaa acatccattc tatcttgtgt aatttttggc atggtgaaaa
     1321 agataatcag ccttataatc tttacaaaag taagaaattc tgtaaataag ccttaatgcc
     1381 cttgctttaa attaaaatgg ttctttttca tgataatgtt tgcactttgt gaatatattt
     1441 tagatagttc tgtgaggtat aattaagatg ttttagagac ttatacaatt ttgtctttat
     1501 aaattcttaa ttgattttac catcccagtt taactatgct tcgtcggcat ctctgcacat
     1561 gtcgtgtttt cttaccgtat tgtcctacca agaacctctt ttttgcttgg atcgaaatta
     1621 aaggtttaaa agcaaagtta tggatgctag agtatttcaa agctattcag ctagagctga
     1681 ggggatgaaa aatcccattg ccaaggaatt gttggctttg atggaagaaa agcaaagcaa
     1741 cttgtcagtc gcggtcgatt tgacgaagaa atccgaaatc ttagaattgg tagataaaat
     1801 tggaccctat gtctgtgtta tcaagacaca tattgacgtt gtcgaggatt tcgaccagga
     1861 tatggtagaa aaactggtgg ccttaggtaa aaagcatcgt tttcttatct ttgaggatcg
     1921 caaattcgca gacattggaa ataccgtcaa gctacaatat gcatctggtg tgtacaaaat
     1981 tgcttcttgg gctcatatca caaattgcca tacagtgcca ggcgagggta ttatacaagg
     2041 cctcaaagaa gttggtttac ctttgggacg tggtctcttg cttttggctg aaatgtcttc
     2101 caaaggctct ttggctactg gttcctacac agagaaaacc ttagaatggt ttgagaagca
     2161 taccgatttt tgctttggct ttatagctgg tcgtcgattt cctaaccttc aaagcgacta
     2221 cataactatg tcccctggta tcggcttgga tgttaaagga gacgggctgg gacagcaata
     2281 tcgtactcct gaagaagtga ttgtaaactg cggtagcgat atcatcattg ttggtcgtgg
     2341 agtttatgga gctggtcgta atcctgttgt cgaagccaag agatatagag aagctggttg
     2401 gaaggcatat cagcaaagac tttctcagca ttaaaaaaag actaatgtaa aatttttttg
     2461 gttggttatt gaaaaagtcg atgccttgtt tgcgtttgtt ttcctaggcg ttttatgtca
     2521 gaaggcattt agaattagta tacaagtact ctttggtaaa attttatgta gcgactaaaa
     2581 tattaactat tatagataaa caccttggga ataaaaagta atttgctata gtaatttatt
     2641 aaacatgctc ctacaacatt accacaatct tttctcttgg attgacattg aataagaaaa
     2701 gagtgaattt ttttagactt gtaatgataa ctatgtacaa agccaatgaa agatgtatgt
     2761 agatgaatgt aaaataccat gtagacaaac aagataaaac ttggttataa acattggtgt
     2821 tggaacagaa taaattagat gtcaaaaagt ttcgtcaata tcacaagctt gctagggcgg
     2881 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     2941 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3001 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3061 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3121 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3181 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3241 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3301 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3361 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3421 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3481 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3541 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggcccgcta gcatgcgatc
     3601 tacgataatg agacggtgtt tgggttgttt ttactgaatt tttcttttat tttatgacat
     3661 cttaattttt atgttaatga ttattcttat acttttatct atctttgatt aaaactaata
     3721 ttcattcttt tgatcactaa ttatttttat agtatacgat tcaacccaac tataatacat
     3781 tgtatcaaga gaaattttta attagatgta atttaatagc catttgataa aaaaaaatta
     3841 cctttttttt tgctagtgtt aaattattga tcataacaat gtcgaatgaa tactaataaa
     3901 atagggacac gcgagttgca aatacttgaa atacctttga agatttgtct gtgaattttg
     3961 ataagatttt tatgaccatc tcttttatat tcgatatcgt gaactttcat tttcttttgt
     4021 tttttgcttt tcaagttaaa taacgcggaa attatatttc cttatatgtt tataagagct
     4081 tttaaaggat ttgctatatc gcttacagta ctacaaccgg ttgctcaatg ttatccgttt
     4141 ccgcggtctt cttccgtgcg cattctgccg tgaagtgcta acgctgtgtg tggctttatc
     4201 aaaaaagtgg ctcaaatata gcaatttcat acatatttaa acacccatta tacttaaatg
     4261 tatattttaa aatatataag agattttcac tgtataactg tgtatcttat ctatatgtaa
     4321 tcaacggctt cataccacct cagccagccg tgttataact taccgtttac caactacatt
     4381 ttttgtaacg aaccaaaaaa ccctcaaaag acaagaccga attcatggca agctccaagg
     4441 gtgaagagct atttactggg gttgtaccca ttttggtaga actggacgga gatgtaaacg
     4501 gacataaatt ctctgttaga ggtgagggcg aaggcgatgc caccaatggt aaattgactc
     4561 tgaagtttat atgcactacg ggtaaattac ctgttccttg gccaacccta gtaacaactt
     4621 tgacatatgg tgttcaatgt ttctcaagat acccagacca tatgaaaagg catgatttct
     4681 ttaaaagtgc tatgccagaa ggctacgtgc aagagagaac tatctccttt aaggatgacg
     4741 gtacgtataa aacacgagca gaagtgaaat tcgaagggga tacactagtt aatcgcatcg
     4801 aattaaaggg tatagacttt aaggaagatg gtaatattct cggccataaa cttgagtata
     4861 atttcaactc gcataatgtg tacattacag ctgacaaaca aaagaacgga attaaagcga
     4921 attttaaaat caggcacaac gtcgaagatg ggtctgttca acttgccgat cattatcagc
     4981 aaaacacccc tattggtgat ggtccagtct tgttacccga taatcactac ttaagcacac
     5041 agtctagatt gtcaaaagat ccgaatgaaa agcgtgatca catggtttta ttggaatttg
     5101 tcaccgctgc aggaataact cacggaatgg acgagcttta taagccccga tgccctttct
     5161 tcgagacgtc cgcagcttac agatattata tagatgacgt attccatgcc cttgtacgcg
     5221 agatacgtcg aaaggaaaag gaagcagttc ttgctatgga aaaaaaatct aaacccaaga
     5281 actcagtgtg gaagagattg aagagtcctt tccgaaaaaa aaaggacagt gtaactctcg
     5341 agtgaggcgc gccataagcg gccgccaccg cggtggagct cggctctaga gcttttatga
     5401 ttttgataaa tacttttttt cgttttggtt tgtgatacgt atagtttgcc tgtctccttt
     5461 tgtaaatgat cttcttttgt agtcttgtgt aatctctttt tcatttggtc aacttaagtg
     5521 aatttttttt tgtcagtttc cttgtaggat attatgtcaa agtcttattt atactatata
     5581 ttattgcttt ggtggtgagt cgcatttgtg gggacggtgt tactttactg atggatttag
     5641 gtaggaggca tccactgtcc cccatttcgc tagcttaagt ttaatatgaa gatgcactta
     5701 agcatgaacg cccgttttga tctttttcac ttgtaccgcg aagagcttgt tttactgttt
     5761 ttccgtctgc atcccgagaa ctagttgatc tcccaatgca cactttagac tttattcaaa
     5821 ccgaagtcgg ttttgtaatt taaaaatttt tacccactgt aaagtacaaa caattcatcg
     5881 ctatgttagt tgtgtacgct actagcgcat cacatgatta catgttaatc aacaaaaacg
     5941 aaaatgttta ttgtcaatct aaaaatacat atgtccgtta agacaaaagt atgaggctgg
     6001 atggaatgag caatggtagc aaaaggtttc cgaaatggtc atatcatttt ttcaattttt
     6061 gctgatctca aatacctcaa cttgttttat tcgtttaaac cgagctccag cttttgttcc
     6121 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     6181 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6241 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6301 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6361 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6421 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6481 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6541 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6601 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6661 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6721 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6781 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6841 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6901 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6961 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7021 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7081 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7141 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7201 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7261 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7321 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7381 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7441 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7501 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7561 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7621 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7681 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7741 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7801 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7861 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7921 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7981 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8041 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8101 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8161 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8281 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8341 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
