LOCUS       sequence-366161-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector to add a triple repeat of the amino acid linker 
            Gly-Gly-Gly-Gly-Ser [(GGGGS)x3]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3007)
  AUTHORS   Shimozono S, Miyawaki A
  TITLE     Engineering FRET constructs using CFP and YFP.
  JOURNAL   Methods Cell Biol. 2008;85:381-93. doi: 
            10.1016/S0091-679X(08)85016-9.
  PUBMED    18155471
REFERENCE   2  (bases 1 to 3007)
  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: "Methods 
            Cell Biol."; date: "2008"; volume: "85:381-93. doi"; pages: " 
            10.1016/S0091-679X(08)85016-9"
COMMENT     Sequence Label: pBS Coupler 2
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..85)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(69..85)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(117..136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(118..136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(146..163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(146..162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(155..177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(303..758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(440..461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     652..671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        785..889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(890..958,959..1750)
                     /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(1108..1127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1921..2509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2410..2429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2663..2680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2797..2818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2833..2838,2839..2856,2857..2863)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2871..2887
                     /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     2876..2898
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2895..2911
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2895..2911
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2930..2950
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2932..2950
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2979..2995
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2980..2996
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcgcatcg aattcctgca
       61 gcccggggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc aattcgccct
      121 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      181 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      241 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      301 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      361 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      421 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      481 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      541 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      601 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      661 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      721 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
      781 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      841 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      901 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      961 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1021 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1081 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1141 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1201 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1261 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1321 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1381 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1441 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1501 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1561 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1621 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1681 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1741 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1801 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1861 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2041 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2581 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2641 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2701 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2761 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2821 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     2881 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     2941 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
     3001 gcttgat
//
