LOCUS       sequence-181158-        3137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 Linker containing between FS1 and FS2 the BB3 linker with FSA 
            and FSB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3137)
  AUTHORS   Sarkari P, Marx H, Blumhoff ML, Mattanovich D, Sauer M, Steiger MG
  TITLE     An efficient tool for metabolic pathway construction and gene 
            integration for Aspergillus niger.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004.
  PUBMED    28533066
REFERENCE   2  (bases 1 to 3137)
  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: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: BB2_L_12_syn_A_B
FEATURES             Location/Qualifiers
     source          1..3137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..64
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    181..202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(217..222,223..240,241..247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    255..271
                     /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     260..282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     279..295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     279..295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     314..333
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        314..332
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(580..598)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(581..598)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(612..629)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(612..628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(621..643)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(769..1224)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(906..927)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1118..1137
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1251..1355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1356..1424,1425..2216)
                     /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(1574..1593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2442..3030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2931..2950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
       61 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      121 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      241 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      301 ttacgccaag ctctaatacg actcactata gggaaagctc ggtaccacgc atgctgcaga
      361 cgcgttacgt atcggatcca gaattcgtga tccgggaaga cgccatgccg gggagaccct
      421 cgagggatcc ggtctccgat cgcgaaaaaa ccccgccctg tcaggggcgg ggttttttgc
      481 gctccgcgtc ttcgatcatc tgaattcgtc gacaagcttc tcgagcctag gctagctcta
      541 gaccacacgt gtgggggccc gagctcgcgg ccgctgtatt ctatagtgtc acctaaatgg
      601 ccgcacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      661 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg
      781 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
      841 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      901 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      961 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1021 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1081 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1141 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1201 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1321 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1381 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1441 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1501 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1561 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1621 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1681 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1741 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1801 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1861 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1921 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1981 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2041 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2101 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2161 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2221 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2281 ggatctaggt gaagatcctt tttgataatc tcatgaacaa taaaactgtc tgcttacata
     2341 aacagtaata caaggggtgt agatctcatg accaaaatcc cttaacgtga gttttcgttc
     2401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2581 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3121 gataaccgta ttaccgc
//
