LOCUS       sequence-366159-        3008 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 3008)
  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 3008)
  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 3
FEATURES             Location/Qualifiers
     source          1..3008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..86)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(70..86)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(118..137)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(119..137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(147..164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(147..163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(156..178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(304..759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(441..462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     653..672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        786..890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(891..959,960..1751)
                     /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(1109..1128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1922..2510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2411..2430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2664..2681
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2798..2819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2834..2839,2840..2857,2858..2864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2872..2888
                     /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     2877..2899
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2896..2912
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2896..2912
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2931..2951
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2933..2951
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2980..2996
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2981..2997
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcgccatc gaattcctgc
       61 agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc caattcgccc
      121 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
      181 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      241 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      301 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
      361 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
      421 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
      481 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
      541 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
      601 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
      661 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
      721 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
      781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      841 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      901 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      961 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1021 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1081 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1141 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1201 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1261 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1321 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1381 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1441 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1501 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1561 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1621 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1681 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1741 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1801 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1861 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1921 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1981 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2041 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2101 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2161 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2221 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2281 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2341 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2401 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2461 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2521 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2581 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2641 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2701 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2761 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     2821 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2881 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     2941 ctcactaaag ggaacaaaag ctgggtaccg ggccccccct cgaggtcgac ggtatcgata
     3001 agcttgat
//
