LOCUS       sequence-72920-e        3950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector at position 2, small linker insert, final 
            entry vector for the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3950)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 3950)
  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 One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGEV_2'_Linker
FEATURES             Location/Qualifiers
     source          1..3950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        971..989
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     971..988
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          1098..1670
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    1743..1877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1767..1786)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1821..1840
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2029..2046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2200..2788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(2959..3750,3751..3819))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3582..3601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3820..3924)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctacgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagggc gcctccccca
      541 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
      601 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
      661 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
      721 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
      781 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
      841 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
      901 tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg gggacgtcgg
      961 agcaagcttg atttaggtga cactatagag gtctcaggct ccggatccgg aggcggaagc
     1021 ggaggaggta cctaagtgag accctcatcc tcccgggtaa ctaagtaagg atctcgaccg
     1081 atcctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     1141 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     1201 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     1261 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     1321 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     1381 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     1441 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     1501 gctggcgtgg aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt
     1561 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     1621 aaccgggccc ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attaattcct cgacggatcg
     1741 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1801 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1861 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1921 aggtgtggga ggttttttaa ttcgcggccg ctctagaact agtctgcatt aatgaatcgg
     1981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tcggcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgttccccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     2401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2641 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3001 tctatttcgt tcatccatag ttgcctgact gcccgtcgtg tagataacta cgatacggga
     3061 gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc
     3121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3361 catgttgtga aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3421 ggccgcagtg ttatcactca tgcttatggc agcactgcat aattctctta ctgtcatgcc
     3481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
