LOCUS       sequence-420105-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian overexpression vector for C-terminally StayGold tagged 
            human Prom1s1 (W795R).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  AUTHORS   Bell TA, Luce BE, Hakim P, Ananda VY, Dardari H, Nguyen TH, 
            Monshizadeh A, Chao LH
  TITLE     Prominin 1 and Tweety Homology 1 both induce extracellular vesicle 
            formation.
  JOURNAL   Elife. 2024 Aug 13;13:e100061. doi: 10.7554/eLife.100061.
  PUBMED    39168849
REFERENCE   2  (bases 1 to 7330)
  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: "Elife."; 
            date: "2024-08-13"; volume: "13:e100061. doi"; pages: " 
            10.7554/eLife.100061"
COMMENT     Sequence Label: pCS2-HsProm1s1(W795R)-StayGold
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2666..2686
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2699..3349
                     /codon_start=1
                     /product="bright and highly photostable dimeric green 
                     fluorescent protein (Hirano et al., 2022)"
                     /label=StayGold
                     /note="engineered variant of a GFP from Cytaeis uchidae"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     polyA_signal    3382..3516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3406..3425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3460..3479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3629..3649)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3629..3647)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3668..3684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3668..3684)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3681..3703)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3692..3708
                     /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(3716..3722,3723..3740,3741..3746))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3761..3782
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3899..3916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4070..4658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4150..4169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4829..5620,5621..5689))
                     /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     5452..5471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5690..5794)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5820..6275)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5957..5978)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6169..6188
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6346..7330
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tatggccctc gtactcggct ccctgttgct
      121 gctggggctg tgcgggaact ccttttcagg agggcagcct tcatccacag atgctcctaa
      181 ggcttggaat tatgaattgc ctgcaacaaa ttatgagacc caagactccc ataaagctgg
      241 acccattggc attctctttg aactagtgca tatctttctc tatgtggtac agccgcgtga
      301 tttcccagaa gatactttga gaaaattctt acagaaggca tatgaatcca aaattgatta
      361 tgacaagatt gtctactatg aagcagggat tattctatgc tgtgtcctgg ggctgctgtt
      421 tattattctg atgcctctgg tggggtattt cttttgtatg tgtcgttgct gtaacaaatg
      481 tggtggagaa atgcaccagc gacagaagga aaatgggccc ttcctgagga aatgctttgc
      541 aatctccctg ttggtgattt gtataataat aagcattggc atcttctatg gttttgtggc
      601 aaatcaccag gtaagaaccc ggatcaaaag gagtcggaaa ctggcagata gcaatttcaa
      661 ggacttgcga actctcttga atgaaactcc agagcaaatc aaatatatat tggcccagta
      721 caacactacc aaggacaagg cgttcacaga tctgaacagt atcaattcag tgctaggagg
      781 cggaattctt gaccgactga gacccaacat catccctgtt cttgatgaga ttaagtccat
      841 ggcaacagcg atcaaggaga ccaaagaggc gttggagaac atgaacagca ccttgaagag
      901 cttgcaccaa caaagtacac agcttagcag cagtctgacc agcgtgaaaa ctagcctgcg
      961 gtcatctctc aatgaccctc tgtgcttggt gcatccatca agtgaaacct gcaacagcat
     1021 cagattgtct ctaagccagc tgaatagcaa ccctgaactg aggcagcttc cacccgtgga
     1081 tgcagaactt gacaacgtta ataacgttct taggacagat ttggatggcc tggtccaaca
     1141 gggctatcaa tcccttaatg atatacctga cagagtacaa cgccaaacca cgactgtcgt
     1201 agcaggtatc aaaagggtct tgaattccat tggttcagat atcgacaatg taactcagcg
     1261 tcttcctatt caggatatac tctcagcatt ctctgtttat gttaataaca ctgaaagtta
     1321 catccacaga aatttaccta cattggaaga gtatgattca tactggtggc tgggtggcct
     1381 ggtcatctgc tctctgctga ccctcatcgt gattttttac tacctgggct tactgtgtgg
     1441 cgtgtgcggc tatgacaggc atgccacccc gaccacccga ggctgtgtct ccaacaccgg
     1501 aggcgtcttc ctcatggttg gagttggatt aagtttcctc ttttgctgga tattgatgat
     1561 cattgtggtt cttacctttg tctttggtgc aaatgtggaa aaactgatct gtgaacctta
     1621 cacgagcaag gaattattcc gggttttgga tacaccctac ttactaaatg aagactggga
     1681 atactatctc tctgggaagc tatttaataa atcaaaaatg aagctcactt ttgaacaagt
     1741 ttacagtgac tgcaaaaaaa atagaggcac ttacggcact cttcacctgc agaacagctt
     1801 caatatcagt gaacatctca acattaatga gcatactgga agcataagca gtgaattgga
     1861 aagtctgaag gtaaatctta atatctttct gttgggtgca gcaggaagaa aaaaccttca
     1921 ggattttgct gcttgtggaa tagacagaat gaattatgac agctacttgg ctcagactgg
     1981 taaatccccc gcaggagtga atcttttatc atttgcatat gatctagaag caaaagcaaa
     2041 cagtttgccc ccaggaaatt tgaggaactc cctgaaaaga gatgcacaaa ctattaaaac
     2101 aattcaccag caacgagtcc ttcctataga acaatcactg agcactctat accaaagcgt
     2161 caagatactt caacgcacag ggaatggatt gttggagaga gtaactagga ttctagcttc
     2221 tctggatttt gctcagaact tcatcacaaa caatacttcc tctgttatta ttgaggaaac
     2281 taagaagtat gggagaacaa taataggata ttttgaacat tatctgcagt ggatcgagtt
     2341 ctctatcagt gagaaagtgg catcgtgcaa acctgtggcc accgctctag atactgctgt
     2401 tgatgtcttt ctgtgtagct acattatcga ccccttgaat ttgtttcgtt ttggcatagg
     2461 aaaagctact gtatttttac ttccggctct aatttttgcg gtaaaactgg ctaagtacta
     2521 tcgtcgaatg gattcggagg acgtgtacga tgatgttgaa actataccca tgaaaaatat
     2581 ggaaaatggt aataatggtt atcataaaga tcatgtatat ggtattcaca atcctgttat
     2641 gacaagccca tcacaacatc tcgaggagaa tttgtacttt caaggcggtg gaggagggat
     2701 ggcctcaaca cccttcaagt tccaattgaa gggcacaatc aacggtaaat ctttcactgt
     2761 ggagggcgag ggggagggca acagccatga agggtctcac aaaggcaaat acgtgtgcac
     2821 gagcgggaag ctgcccatgt catgggccgc tctggggaca agctttgggt atggaatgaa
     2881 atactacact aagtatccta gtgggcttaa gaactggttt cacgaggtga tgcctgaagg
     2941 cttcacatac gataggcata ttcagtacaa aggcgacgga agtatccatg caaagcacca
     3001 acattttatg aagaatggga cctatcataa catcgtggaa tttactggcc aggactttaa
     3061 ggaaaacagc cccgtactga caggagacat gaatgtcagt ctccctaacg aggtgcagca
     3121 tatcccccga gacgacggcg tggagtgtcc cgtgactctc ctgtacccgc tcctctccga
     3181 taagtccaag tgtgtggagg cccaccaaaa tactatctgt aaacctctgc ataatcagcc
     3241 cgctcccgat gtaccatacc attggattcg gaagcagtac acccagagca aggatgacac
     3301 cgaagagcgg gatcatatat gtcaaagcga aaccctcgag gcccacctct aatctagaac
     3361 tatagtgagt cgtattacgt agatccagac atgataagat acattgatga gtttggacaa
     3421 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3481 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3541 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg
     3601 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     3661 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3721 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3781 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4261 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4501 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4621 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4681 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4741 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4801 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4921 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4981 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5101 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5161 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5221 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5281 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5341 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5401 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5461 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5521 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5581 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5641 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5701 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5761 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5821 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5881 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5941 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6001 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6061 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6121 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6181 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6241 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     6301 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccgccaattc
     6361 aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc
     6421 aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc
     6481 aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact gtgccaactg
     6541 gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt
     6601 ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat atgccaggat
     6661 caatatatag gcaatatcca atatggccct atgccaatat ggctattggc caggttcaat
     6721 actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta ttgacgtaga
     6781 tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat accgcccata
     6841 gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg acgtcatatg
     6901 ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg taaatggccc
     6961 gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc aatgggatgg
     7021 ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac ggtaaatggc
     7081 ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt actattggaa
     7141 gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa tggcccgcga
     7201 tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc aaatgggcgt
     7261 tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg
     7321 tttagggaac
//
