LOCUS       sequence-454235-        8691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry plasmid of hBIG1 for Gateway system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8691)
  TITLE     Mammalian expression plasmids and entry plasmids for Gateway
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8691)
  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            "Unpublished"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: pENTR/hBIG1
FEATURES             Location/Qualifiers
     source          1..8691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      88..174
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      266..293
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     301..324
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    343..442
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     mobile_element  2573..3340
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     primer_bind     complement(6867..6883)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6867..6883)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(6905..7004)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7116..7135)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7127..7936
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7194..7213)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8026..8614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8515..8534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc tagcatggat ctcggggacg tctaactact
       61 aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
      121 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      181 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
      241 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
      301 ctacaaactc ttcctgttag ttagttactt aagctcgggc cccaaataat gattttattt
      361 tgactgatag tgacctgttc gttgcaacaa attgataagc aatgcttttt tataatgcca
      421 actttgtaca aaaaagcagg ctctttaaag gaaccaattc agtcgactgg atccaccatg
      481 tatgagggga agaagacgaa gaacatgttc ctgacccggg ctctggagaa gatattggcc
      541 gacaaggaag tgaagaaggc gcatcactcc cagctgcgca aagcttgcga ggtggcgtta
      601 gaggaaataa aagcggaaac tgaaaaacag agtcctcctc atggagaagc aaaagctgga
      661 tcaagcaccc ttccaccagt gaaatcaaag acaaatttta ttgaagcaga caagtacttc
      721 ttgccttttg agttggcatg ccagtccaaa tgtcctcgca tagttagtac atctctagat
      781 tgcttacaga aacttattgc ttatgggcac ttgactggca atgctccaga tagtacaaca
      841 ccaggcaaaa aattaattga tagaattatt gaaacaatat gtggctgctt tcagggccct
      901 cagacagatg aaggagttca gctgcagata ataaaggctt tacttactgc agtaacatca
      961 caacacatag aaattcatga agggactgta ctgcaagctg tgagaacatg ttacaatatc
     1021 tacttagcaa gcaaaaatct catcaatcag acaacagcca aagctactct cactcagatg
     1081 ctaaatgtta tctttgcacg catggaaaac caagcattac aggaagcaaa acaaatggaa
     1141 aaagaaaggc atcggcagca tcatcatctg ttacagtctc cagtaagcca tcacgagcct
     1201 gaatcacctc aacttagata tttgccacct cagactgttg atcatatatc ccaagaacac
     1261 gaaggggacc ttgacctcca tacaaatgat gtagataaaa gtcttcagga tgacacagag
     1321 cctgaaaatg gatctgatat ttccagtgca gaaaatgaac agactgaagc tgatcaggca
     1381 actgcagctg aaacattatc taaaaatgaa gtgttatatg acggagaaaa ccatgattgt
     1441 gaggaaaagc cacaagacat tgtacagaac attgtagaag aaatggtgaa cattgttgtt
     1501 ggagatatgg gagaagggac tactataaat gcaagtgcag atggcaacat tggaactata
     1561 gaggatggta gtgacagtga aaatattcaa gcaaatggaa ttccaggaac accaatttct
     1621 gttgcatata caccatcctt acctgatgat agattgtcag tctcttccaa tgatactcag
     1681 gaatctggaa attcttcagg accttcacct ggtgctaagt tttcccacat tttacaaaag
     1741 gatgcctttc tagtattcag gtcattgtgt aaactgtcaa tgaaaccact gtcagatgga
     1801 ccaccagatc caaagtctca tgaactacga tccaagattc tttcattgca gttacttcta
     1861 tccattctgc agaatgcagg acctattttc aggacaaatg agatgtttat taatgctatt
     1921 aagcagtatc tttgtgttgc actctcaaaa aatggagtct catctgttcc agaggttttt
     1981 gagctttctc tttctatatt tcttactttg ttgtcaaatt tcaagacaca tctgaagatg
     2041 caaattgagg tgttctttaa agaaattttc ttatacattt tggaaacttc taccagctca
     2101 tttgatcaca aatggatggt tattcagaca ctgacgagga tttgtgcaga tgctcagagt
     2161 gtagtggata tttatgtaaa ctatgactgt gacttaaatg cagccaatat atttgaaaga
     2221 ctagtaaatg atctatcaaa aattgctcaa ggaaggggca gtcaagaact tggtatgagt
     2281 aatgttcagg aattgagcct gaggaaaaaa ggtttagaat gcttagtgtc gattttgaag
     2341 tgtatggttg aatggagtaa ggatcagtat gtgaatccca actcccagac aactcttggt
     2401 caggaaaaac cctcagagca agagatgagt gaaatcaaac accctgagac aataaacaga
     2461 tacggaagtt taaattccct ggagtcaaca tcatcatcag gaataggcag ctacagtaca
     2521 cagatgtctg gcactgataa tccagaacaa tttgaggtcc taaagcaaca aaggtaatga
     2581 ctccaactta ttgatagtgt tttatgttca gataatgccc gatgactttg tcatgcagct
     2641 ccaccgattt tgagaacgac agcgacttcc gtcccagccg tgccaggtgc tgcctcagat
     2701 tcaggttatg ccgctcaatt cgctgcgtat atcgcttgct gattacgtgc agctttccct
     2761 tcaggcggga ttcatacagc ggccagccat ccgtcatcca tatcaccacg tcaaagggtg
     2821 acagcaggct cataagacgc cccagcgtcg ccatagtgcg ttcaccgaat acgtgcgcaa
     2881 caaccgtctt ccggagactg tcatacgcgt aaaacagcca gcgctggcgc gatttagccc
     2941 cgacatagcc ccactgttcg tccatttccg cgcagacgat gacgtcactg cccggctgta
     3001 tgcgcgaggt taccgactgc ggcctgagtt ttttaagtga cgtaaaatcg tgttgaggcc
     3061 aacgcccata atgcgggctg ttgcccggca tccaacgcca ttcatggcca tatcaatgat
     3121 tttctggtgc gtaccgggtt gagaagcggt gtaagtgaac tgcagttgcc atgttttacg
     3181 gcagtgagag cagagatagc gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc
     3241 gtcagtagct gaacaggagg gacagctgat agaaacagaa gccactggag cacctcaaaa
     3301 acaccatcat acactaaatc agtaagttgg cagcatcacc agcaacaaaa agaaataata
     3361 gaacaaggga tagatttatt taataagaaa ccaaagagag gaatacagta cctccaagaa
     3421 caagggatgc ttggcaccac acctgaagat attgcccaat tcttacatca agaggaaaga
     3481 ttagactcta ctcaagtggg tgagttcctg ggagataatg ataaatttaa caaagaagtc
     3541 atgtatgcat atgtggacca acatgacttt tcaggaaaag acttcgtttc agcccttcgt
     3601 atgtttctag aaggatttcg tcttccaggg gaagctcaga aaatcgatcg attaatggaa
     3661 aaatttgctg caagatacct agaatgcaac caaggacaaa ctctctttgc tagtgcggat
     3721 acagcttatg ttttggctta ttcaattatc atgttgacca cagaccttca cagtccacag
     3781 gtgaaaaata aaatgacaaa ggaacaatac attaagatga atagaggtat caatgacagt
     3841 aaagaccttc ctgaagagta tctatcagcc atctataatg aaatagctgg gaaaaagata
     3901 tcaatgaaag aaacaaaaga actaacaatc cctacaaaat caagtaaaca gaatgtagcc
     3961 agtgaaaaac aaagaagact tctgtataac ttagaaatgg agcagatggc caagacagct
     4021 aaagcactca tggaggccgt gagccatgtt caggcaccct ttacaagtgc aacacatttg
     4081 gagcatgtga ggcccatgtt taagttggct tggacgcctt ttctggctgc attcagtgtg
     4141 ggtctacaag attgtgatga tactgaagta gcctctcttt gcctggaagg tataagatgt
     4201 gcaatcagaa ttgcatgcat tttcagcatt cagctggaga gagatgcata tgtccaggca
     4261 ctagcaagat ttaccttact cacagtgagt tccggtatta ctgaaatgaa acagaagaac
     4321 attgacacaa taaaaacact tatcacagtg gctcatacag atggaaatta tttaggaaat
     4381 tcatggcatg agattctgaa gtgcatcagt cagttggagc tggcacagct tataggaact
     4441 ggagtgaaac ctcgatacat ttctggaaca gtgcgaggca gagaaggatc tcttactgga
     4501 acaaaagatc aggctcctga tgaatttgtg ggtttagggc tagttggagg aaatgtggac
     4561 tggaaacaga tagcaagtat tcaggaatcc attggagaaa ccagttctca gagtgttgtt
     4621 gttgctgtag atagaatatt cacaggatct acaaggctag atggaaatgc cattgtggat
     4681 tttgtccgtt ggctctgtgc tgtgtctatg gatgaattac tttccacgac acacccaaga
     4741 atgtttagtc tacaaaaaat agtagaaata tcatattaca acatgggaag aataagatta
     4801 cagtggtctc gaatttggga agttattgga gatcatttta ataaggttgg gtgtaatcct
     4861 aatgaagatg tagctatttt tgcagtagac tccttgaggc agttgtcaat gaagttctta
     4921 gagaaagggg agcttgctaa cttcagattc cagaaggatt tcttaagacc ttttgaacat
     4981 ataatgaaac ggaacaggtc tccaacaatt cgagatatgg ttgtacggtg tatagcacag
     5041 atggttaatt ctcaagctgc taacattcga tctggatgga agaacatttt ctctgtattt
     5101 catctagctg catctgatca agatgaaagc atagtggaac ttgcattcca aacaaccggg
     5161 cacattgtca cccttgtatt tgaaaaacac tttccagcga ccattgattc tttccaggat
     5221 gcagtgaagt gtttgtctga atttgcgtgc aatgcagctt tcccagacac aagtatggaa
     5281 gcaattcgac ttattcgcca ttgtgcaaaa tatgtgtctg atagacctca ggctttcaag
     5341 gaatacacaa gcgatgatat gaacgtagca cctgaagaca gggtgtgggt gagaggatgg
     5401 ttcccaattc tctttgagtt atcctgtatc atcaatagat gcaaattaga tgtaagaacc
     5461 aggggtttaa cagtaatgtt tgaaataatg aaaacatatg gccacactta tgagaaacac
     5521 tggtggcagg atttatttag aattgttttc agaatctttg acaatatgaa attgccagaa
     5581 caacagacag agaaagctga atggatgaca acaacttgca atcatgcact ttatgcaatc
     5641 tgtgatgtat tcactcagta tttagaagta ctcagtgatg tacttttgga tgacattttt
     5701 gctcagctct actggtgtgt gcagcaagac aatgagcagt tagcgcgatc tggtacaaac
     5761 tgtttagaga atgttgttat tctgaatggt gaaaaattta ccctagaaat ctgggataaa
     5821 acttgcaact gcacactgga tatcttcaaa accacaatcc cacatgcgct gttgacctgg
     5881 cgacccaatt ctggagaaac tgccccccca cctccatctc ctgtaagtga aaagccattg
     5941 gatacaatat cacagaagtc tgtagatatt catgattcta ttcaaccaag gtctgtggat
     6001 aacagaccac aagcaccact ggtttctgcg tctgctgtta atgaagaagt cagcaaaatt
     6061 aaatctacag caaaatttcc agaacaaaaa ttgtttgctg ccctgttgat taaatgtgtt
     6121 gtgcagctgg aactcatcca gactatcgac aacattgtct tcttcccagc cacaagtaag
     6181 aaagaagatg cagaaaactt agctgcagca cagagagatg cggtggactt tgatgttcgc
     6241 gttgatactc aagaccaagg aatgtaccgc tttttaacat cacaacaact ttttaagcta
     6301 ctggactgct tattagagtc acatagattt gcaaaagcgt ttaattccaa caacgaacag
     6361 aggactgccc tgtggaaagc aggattcaaa ggcaaatcca agcccaacct tctgaagcag
     6421 gagaccagca gcctggcctg tgggctgcgc attctcttcc ggatgtacat ggatgagagc
     6481 cgcgttagtg cctgggagga ggtccagcag aggcttttga atgtccgcag tgaagcacta
     6541 agttacttcc tcactctaac atcagaaagt catcgagaag cctggactaa cttactgctt
     6601 ttgtttctaa ctaaagttct aaagataagt gataataggt ttaaagctca tgcatcattc
     6661 tactaccctc tcttatgtga aattatgcaa tttgacttga ttcctgaact tcgtgctgtt
     6721 cttagaagat tttttctgcg aatcggagta gtttttcaga tatcacaacc acctgaacag
     6781 gaacttggaa taaacaagca atgatgggaa cttaatattt ttgttggcat ttacatccct
     6841 ctgcatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg ctcgagatat
     6901 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     6961 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct
     7021 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     7081 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     7141 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     7201 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     7261 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     7321 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     7381 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     7441 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     7501 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     7561 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     7621 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     7681 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     7741 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     7801 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     7861 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     7921 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag attgggcccc
     7981 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8041 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8101 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8161 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8221 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8281 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     8341 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8401 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8461 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8521 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8581 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8641 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat c
//
