LOCUS       sequence-343481-        3295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HR donor vector for ACTB gene with the revised marker sequence for 
            ASHRA. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3295)
  TITLE     HR donor vector for ACTB gene with the revised marker sequence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3295)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBS-ACTB-C200-GFPfr2
FEATURES             Location/Qualifiers
     source          1..3295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(24..479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(161..182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     373..392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     606..628
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     620..637
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     621..637
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     647..666
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        647..665
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1128..1148)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1128..1146)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1167..1183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1167..1183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1180..1202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1191..1207
                     /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(1215..1221,1222..1239,1240..1245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1260..1281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1398..1415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1569..2157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1649..1668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2328..3119,3120..3188))
                     /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     2951..2970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3189..3293)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
       61 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
      121 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
      181 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
      241 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
      301 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
      361 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
      421 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
      481 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      541 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      601 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
      661 atagggcgaa ttgggtaccg ggccccccct cgagtggtgt gtggggagct gtcacatcca
      721 gggtcctcac tgcctgtccc cttccctcct cagatcattg ctcctcctga gcgcaagtac
      781 tccgtgtgga tcggcggctc catcctggcc tcgctgtcca ccttccagca gatgtggatc
      841 agcaagcagg agtatgacga gtccggcccc tccatcgtcc accttcaaga tccgccacaa
      901 cgttagttag ttgcgttaca ccctttcttg acaaaaccta acttgcgcag aaaacaagat
      961 gagattggca tggctttatt tgtttttttt gttttgtttt ggtttttttt ttttttttgg
     1021 cttgactcag gatttaaaaa ctggaacggt gaaggtgaca gcagtcggtt ggagcgagca
     1081 tcccccaaag tgcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
     1141 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1381 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgca
//
