LOCUS       sequence-347136-        3885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4x SV40 polyA sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3885)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     Additional gene targeting vector construction plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3885)
  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            "Unpublished"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-4xpA
FEATURES             Location/Qualifiers
     source          1..3885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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)."
     primer_bind     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        367..385
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    533..654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    766..887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    999..1120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1232..1353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1398..1414)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1399..1415)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1439..1458)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1440..1458)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1465..1482)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1465..1481)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1474..1496)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1623..2078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1760..1781)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1972..1991
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2105..2209
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2210..2278,2279..3070)
                     /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     complement(2428..2447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3241..3829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3730..3749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gctcgaaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg
      421 ctctagaatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
      481 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
      541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
      601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaactaga
      661 atcataatca gccwtaccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
      721 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
      781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaact agaatcataa
      901 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
      961 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1021 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1081 attctagttg tggtttgtcc aaactcatca atgtatctta actagaatca taatcagcca
     1141 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1201 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     1261 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1321 ttgtggtttg tccaaactca tcaatgtatc ttaactagtg gatcccccgg gctgcaggaa
     1381 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccaattcgcc
     1441 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1501 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1561 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1621 gcaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1681 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     1741 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1801 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1861 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1921 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1981 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2041 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2161 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2221 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2281 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2341 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2401 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2461 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2521 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2581 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2641 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2701 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2761 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2821 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2881 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2941 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3001 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3061 gcattggtaa ctgtcagacc aagtttactc atatatactt tagaytgatt taaaacttca
     3121 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctca
//
