LOCUS       sequence-263741-        8304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP targeted to the endoplasmic reticulum with a 
            signal sequence and a retention signal (AHDL).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8304)
  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 8304)
  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: pAV0773
FEATURES             Location/Qualifiers
     source          1..8304
                     /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(2526..2981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2875..2894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3107..3129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3121..3138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3122..3138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3148..3167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3148..3166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5043..5386
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      5785..5982
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6044..6064)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6044..6062)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6083..6099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6083..6099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6096..6118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6107..6123
                     /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(6131..6137,6138..6155,6156..6161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6314..6331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6485..7073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6565..6584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7244..8035,8036..8104))
                     /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     7867..7886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8105..8209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8277..8295
                     /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 gggccttacg aagttaaatt ataggcacag acatctttgg cattggaaag
      241 atttagtaac aatcataatg tagcacttct taaaaaatct aattttagaa caaaaaggct
      301 taaaaaagaa tggttccgct cttttaaaaa atagtaaaat gtacaatagc aaggttttcg
      361 taggtcgacg gacataacaa ggggaaagag aaataaagca tgtaaacgtc aataaaacat
      421 ctttgattct ggatcaatca tatgccaaat gaagaaaaat agcatttata acaacatgaa
      481 accatatttg agacctttta acgatgaaaa tttattaaat aaaagataaa aaagaaagaa
      541 accgttcaag ttcaatgaaa atttcgtcgt gaagtagaaa ccaagataat ggaaaggctt
      601 cgcaaagcca caaattttcc aatagtcatt ataactgaag attaaggaca gtatactagt
      661 aaaaatcaag tgaattgtgc ctaggatagg tagctaacgt gactgccact gaacctgtgt
      721 agcgtattag atccatcaag atcccttctg ctcatatcat tgctgatctg taaattgacc
      781 gcatcaaatt tgatctcatt tatatcgaga ttgatctata ttattttgcc aatttgcagt
      841 atataaactt atttgattaa gtttttggtc tccttcgcca cgtcttcatt ttgttagttt
      901 aaattttttt gactaactta gaaagggaag tttttaaaaa cgttgaaata ctttttcgat
      961 ttaaaaatat atatttaatt aattttctgt tccaaaatgg ttgctcctca tctttggtta
     1021 cgtgctgaaa ccaaaccttt agaagagcgc agtgctttga ctcctagaac tgccaagata
     1081 ctcgccgatg ctggatttca aattaccatt gagcgctctt cccaacgtgc tttcaaggac
     1141 aaagaatttg agcgccttgg tttccccatg gttcctgagg gctcatggcg acatgcacct
     1201 aaggacgcgt acattattgg tctcaaagag cttccggaaa atgataacag tcctttgaag
     1261 catactcata ttcaatttgc tcattgctac aaaaaccagg aaggttggcg tgaagtcttg
     1321 tcccgattcc ccgctggcaa tggtcttcta tatgatttgg aatttttgca agatgacaat
     1381 ggcagacgtg ttgccgcttt tggttaccac gctggttttg ctggatcagc tatctcatgt
     1441 ttagtttggg cacatcaatt attgcatcct aacaaacaat tccctgctat ccgtccgttc
     1501 cccaacgaaa aatctttggt tcgccatgtt gctcgtcaag tacgtttagc tttgaaaaag
     1561 aataacaacc agtatcctcg tattctggtt attggggctc ttggtcgttg tggtactgga
     1621 gcgtgcgatt tagcatcaaa gattggtatt cccttcgata acattcttcg ctgggacatt
     1681 aatgaaacta agaagggtgg tccctttacc gaaattacgg aatcagacat atttgttaac
     1741 tgcatttatc tgagcatgcc tattcccaaa ttctgtaccg tcgagtcgtt aaatgtacct
     1801 aaccgtaaat tgcgtgttgt ttgtgatgtt agctgtgaca ctaccaatcc taacaatcct
     1861 attccaattt ataatgtaaa cactacattt gatcatccta cagttgaggt taaaggtgtc
     1921 acaactcctc ctcctttaga ggtcatttct attgatcatc ttcctacttt attgccaagg
     1981 gaatcttccg aagcatttag tgaagctcta attcctagtc ttttggcttt gaaggacgtc
     2041 gacaatgctc ccgtttgggt cagagctaaa aaactttatg aaaccatggt tcaaaagctt
     2101 taaaataaat gttatttaat ttcaaagttc cgggtccttt atattgtttc accaaattat
     2161 tgaatattat tttctcaaaa attaacttta gaatacaatt tagacggcat tttcgatgct
     2221 ctggacattt tttccacgta cttgttaaat tgtagcggtg ttgcataata ctaccttttg
     2281 atttgttatt cacgttacaa tatatatttg actaatttaa atatgataat atacacattt
     2341 ctctgcaaac ctcctttata cttttcgccg gtcattgatt aataattaag aaacaggatt
     2401 tcatcctgca tttgccactt acctatatgg taaatataaa tcttgaaact ttccccaata
     2461 tttattgctt gctagggcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2521 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     3001 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     3061 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3121 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     3181 gtgccaaggg aagggtaggt gtcgttaagg tctgaggttt ggtgtcagtg ggagatctaa
     3241 aatattatgg caaaattttc ggcgtcgcaa caataatagt aaacagatgt aaggactata
     3301 ccattttgag atgttaaaat ggtgagcaat gatacaagtg atgatctatc atcttttgct
     3361 gattgataaa aaaagacaaa ccctttaatt atttttagct acataatttt ctcttaatgg
     3421 attaatttgg aaagtgaacg caaaatatta tttagtctaa acatgatcgt atttgagatg
     3481 atgccttttg caagtatgtt atactttatt taaccatgac ttttacaatt caaccaatgc
     3541 gacactatcg gacagttgaa gccaatttta attattagct atatatttta gcgtttttcc
     3601 attatatttg tagttaaaac agctatttat aggccattat caaatatata tataattttt
     3661 tttttttttt tttttctctt tcatttggga atatagtgaa ataactctat ataacacaat
     3721 atcaaaataa agatttccca ccttcactgc ttttgttctt ttcacaatta ttcatttcag
     3781 aatttaccaa tttcgatttt cattgtttac attcattgct atgaaaacgt aaggtggtgg
     3841 cgcaatagga cttatcgaaa tgtacagaac tcactataga attgttgtgt tgatgagctt
     3901 caactgcatt cttctggaaa gtactagtat taacgacgtg actgctcctc tcgttactta
     3961 gctgatttct ggtacgctat taaactcatc caaaaccaac tattctagtt tggtaaatct
     4021 taatcaaaaa ctattaaaac ccgtttacta tttacttaac aggttgtttt caataattgg
     4081 gaattggctt gtgcctacga tctcttgtaa ttgaactaca catataagca tttataagtt
     4141 ggtaatcttc aaattcttgt ttattgaaaa tgaagaagtt ccagctattt agcattttaa
     4201 gctactttgt agctttattc ctcctaccta tggcttttgc tagtggatcc ggtggtcccg
     4261 gtggtatgtc caagggtgaa gagctattta ctggggttgt acccattttg gtagaactgg
     4321 acggagatgt aaacggacat aaattctctg ttagaggtga gggcgaaggc gatgccacca
     4381 atggtaaatt gactctgaag tttatatgca ctacgggtaa attacctgtt ccttggccaa
     4441 ccctagtaac aactttgaca tatggtgttc aatgtttctc aagataccca gaccatatga
     4501 aaaggcatga tttctttaaa agtgctatgc cagaaggcta cgtgcaagag agaactatct
     4561 cctttaagga tgacggtacg tataaaacac gagcagaagt gaaattcgaa ggggatacac
     4621 tagttaatcg catcgaatta aagggtatag actttaagga agatggtaat attctcggcc
     4681 ataaacttga gtataatttc aactcgcata atgtgtacat tacagctgac aaacaaaaga
     4741 acggaattaa agcgaatttt aaaatcaggc acaacgtcga agatgggtct gttcaacttg
     4801 ccgatcatta tcagcaaaac acccctattg gtgatggtcc agtcttgtta cccgataatc
     4861 actacttaag cacacagtct agattgtcaa aagatccgaa tgaaaagcgt gatcacatgg
     4921 ttttattgga atttgtcacc gctgcaggaa taactcacgg aatggacgag ctttataagg
     4981 aggccgatga actttaagag ctcgtttagc ttgcctcgtc cccgccgggt cacccggcca
     5041 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     5101 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     5161 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     5221 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     5281 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     5341 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgc ctttgtctca
     5401 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     5461 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     5521 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     5581 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     5641 gagcccctgc ggacggtgtc gacaggtgct tctcgatctg catcctggga tcaaagccat
     5701 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     5761 ttatgtgtgg gagggctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     5821 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     5881 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     5941 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     6001 cgccgccatc cacctgcagg taatcacgtg ccagcttttg ttccctttag tgagggttaa
     6061 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6121 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6181 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6241 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8281 ggcgtatcac gaggcccttt cgtc
//
