LOCUS       sequence-420108-        7339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian overexpression vector for C-terminally mScarlet tagged 
            human Prom1s1 (WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7339)
  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 7339)
  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-mScarlet
FEATURES             Location/Qualifiers
     source          1..7339
                     /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"
     polyA_signal    3391..3525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3415..3434)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3469..3488
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3638..3658)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3638..3656)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3677..3693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3677..3693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3690..3712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3701..3717
                     /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(3725..3731,3732..3749,3750..3755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3770..3791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3908..3925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4079..4667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4159..4178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4838..5629,5630..5698))
                     /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     5461..5480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5699..5803)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5829..6284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5966..5987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6178..6197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6355..7339
                     /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 caaagtacgc 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 ttgttttggt 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 tcgaggtcag caaaggggaa gccgtcatta aagaattcat
     2701 gcgcttcaaa gtccatatgg agggatccat gaacggccac gagttcgaga tcgagggtga
     2761 gggggagggc cggccctacg aggggacaca aactgccaag ctgaaagtga caaagggcgg
     2821 accattgcct ttttcttggg atatcttgag cccccagttc atgtacgggt caagggcatt
     2881 tactaagcac ccggctgata tccctgatta ctataagcag tctttcccag aaggcttcaa
     2941 atgggaacgg gtcatgaact ttgaggacgg cggcgcggtg accgtcactc aggacaccag
     3001 ccttgaagat ggtacactca tttacaaggt taagctgaga ggcacaaact tcccacctga
     3061 tgggccggtg atgcaaaaaa agaccatggg ctgggaggcc agcaccgaga ggctctatcc
     3121 cgaagatggg gtcctcaaag gagatatcaa aatggctctg cgcctcaaag atggcggcag
     3181 gtatctcgca gacttcaaaa caacctacaa ggccaagaaa cccgtccaga tgcccggcgc
     3241 ttacaacgtt gatcgaaaat tggatattac cagtcacaac gaggattaca ctgtggtgga
     3301 gcagtacgaa agaagcgagg gaaggcatag cactggtggg atggacgagc tgtacaaata
     3361 atctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     3421 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3481 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3541 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
     3601 gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     3661 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3721 cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3781 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3841 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3901 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3961 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5821 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5881 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     5941 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6001 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6061 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6121 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6181 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     6241 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     6301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
     6361 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     6421 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     6481 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
     6541 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     6601 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     6661 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     6721 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     6781 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     6841 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     6901 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     6961 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     7021 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     7081 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     7141 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     7201 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     7261 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     7321 caatgctcgt ttagggaac
//
