LOCUS       sequence-150761-        5293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generates B2 inserts suitable for rescue.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5293)
  AUTHORS   Wagstaff BJ, Hedges DJ, Derbes RS, Campos Sanchez R, Chiaromonte F, 
            Makova KD, Roy-Engel AM
  TITLE     Rescuing Alu: recovery of new inserts shows LINE-1 preserves Alu 
            activity through A-tail expansion.
  JOURNAL   PLoS Genet. 2012;8(8):e1002842. doi: 10.1371/journal.pgen.1002842. 
            Epub 2012 Aug 9.
  PUBMED    22912586
REFERENCE   2  (bases 1 to 5293)
  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: "PLoS 
            Genet."; date: "2012"; volume: "8(8):e1002842. doi"; pages: " 
            10.1371/journal.pgen.1002842. Epub 2012 Aug 9"
COMMENT     Sequence Label: pBS B2rescue A70D
FEATURES             Location/Qualifiers
     source          1..5293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1051..1070)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2074..2093
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2174..2221)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2269..2598)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2283..2418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2583..2603
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3059..3075)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3076)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3105..3125)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3105..3123)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3144..3160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3144..3160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3157..3179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3168..3184
                     /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(3192..3198,3199..3216,3217..3222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3237..3258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3375..3392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3546..4134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3626..3645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4305..5096,5097..5165))
                     /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     4928..4947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5166..5270)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccccc tccgctcccc aatgacgtaa
      721 ctgccctgca gcttctagta gcttttcgca gcgtctccga ccgggctggt gagatggctc
      781 agtgggttag agcacccgac tgctcttccg aaggtccaga gttcaaattc cagcaaccac
      841 atggtggctc acaaccatcc gtaacaagat ctgactccct cttctggagt gtctgaagac
      901 agctacagtg tacttacata taagacgtct caaatcccct cagaagaact cgtcaagaag
      961 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     1021 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     1081 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattctc
     1141 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     1201 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     1261 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     1321 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     1381 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     1441 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     1501 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgtaatta
     1561 gcaatataat gaattgcagg gaaaagttat caggcatgca cctggtagct agtctttaaa
     1621 ccaatagatt gcatcggttt aaaaggcaag accgtcaaat tgcgggaaag gggtcaacag
     1681 ccgttcagta ccaagtctca ggggaaactt tgagatggcc ttgcaaaggg tatggtaata
     1741 agctgacgga catggtccta accacgcagc caagtcctaa gtcaacagat cttctgttga
     1801 tatggatgca gttcacagac taaatgtcgg tcggggaaga tgtattcttc tcataagata
     1861 tagtcggacc tctccttaat gggagctagc ggatgaagtg atgcaacact ggagccgctg
     1921 ggacctaatt tgtatgcgaa agtatattga ttaggtttgg agtactcgag ttcattcagg
     1981 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     2041 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     2101 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatggccga tcccatattg
     2161 gctcgacctc cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     2221 cacgtgctga tcagatccga aaatggatat acaagctccc gggagcttta tgcaaaagcc
     2281 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     2341 ggcctctgca taaataaaaa aaattagtca gccatggggc ggaaaatggg cggaactggg
     2401 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     2461 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     2521 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     2581 acaccctaac tgacacacat tccacagcca gatccctggc ttgttgcaca accgttaaac
     2641 cttaaaacct ttaaaagcct tatatattct tgttcttctt ataaaactta aaaccttaga
     2701 ggctatttaa gttgctgatt tatattaata ttattgttca aacatgagag cttagtacgt
     2761 gaaaccatga gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt
     2821 agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga
     2881 gagcttagta cgtactatca acaggttgaa ctgccgtctc aaaaaaaaaa aaaaaaacat
     2941 tacaaaaaaa aaaaaaaaaa agaaaaaaaa aaaaaaaaac acacaaaaaa aaaaaaaaaa
     3001 aacaaggtaa ctggcacaca caaccttttc attttcagga attcgatatc aagcttatcg
     3061 ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta
     3121 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3181 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3241 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3301 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3361 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3721 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3961 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4201 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4261 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4321 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4381 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4441 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4501 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4561 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4621 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4681 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4741 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4801 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4861 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4921 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4981 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5041 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5101 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5161 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5221 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5281 cgaaaagtgc cac
//
