LOCUS       sequence-350392-        2960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone vector for STREAMING-tag knockin vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2960)
  AUTHORS   Ohishi H, Shimada S, Uchino S, Li J, Sato Y, Shintani M, Owada H, 
            Ohkawa Y, Pertsinidis A, Yamamoto T, Kimura H, Ochiai H
  TITLE     STREAMING-tag system reveals spatiotemporal relationships between 
            transcriptional regulatory factors and transcriptional activity.
  JOURNAL   Nat Commun. 2022 Dec 20;13(1):7672. doi: 10.1038/s41467-022-35286-2.
  PUBMED    36539402
REFERENCE   2  (bases 1 to 2960)
  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: "Nat 
            Commun."; date: "2022-12-20"; volume: "13(1):7672. doi"; pages: " 
            10.1038/s41467-022-35286-2"
COMMENT     Sequence Label: pBSK-Delta-B
FEATURES             Location/Qualifiers
     source          1..2960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(18..446)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(128..149)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(229..804)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     340..359
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     573..595
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     587..604
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     588..604
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     614..633
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        614..632
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    641..748
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     657..673
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     658..674
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(708..724)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(708..724)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(761..781)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(761..779)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(800..816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(800..816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(813..835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    824..840
                     /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(848..854,855..872,873..878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    893..914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1031..1048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1202..1790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1282..1301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1961..2752,2753..2821))
                     /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     2584..2603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2822..2912)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
       61 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
      121 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      181 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
      241 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
      301 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
      361 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
      421 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
      481 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      541 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      601 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga
      661 ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat ccactagttc
      721 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
      781 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
      841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2101 ggctgccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2821 tactcttcct tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2881 catatttgaa tgtatttaga aaaataaaca aatagggatt ccgcgcacat ttccccgaaa
     2941 agtgccacct aaattgtaag
//
