LOCUS       sequence-366162-        3006 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 3006)
  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 3006)
  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 4
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(68..84)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(68..84)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(116..135)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(117..135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(145..162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(145..161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(154..176)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(302..757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(439..460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     651..670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        784..888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(889..957,958..1749)
                     /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(1107..1126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1920..2508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2409..2428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2662..2679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2796..2817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2832..2837,2838..2855,2856..2862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2870..2886
                     /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     2875..2897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2894..2910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2894..2910
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2906..3006,1..310)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLIDPPPPEPPPPE
                     PPPPSNSCSPGDPLVLERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     primer_bind     2929..2949
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2931..2949
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2978..2994
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2979..2995
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgatccgcca ccgccagagc cacctccgcc tgaaccgcct ccaccatcga attcctgcag
       61 cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca attcgcccta
      121 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
      181 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      241 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      301 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
      361 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
      421 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
      481 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
      541 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
      601 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
      661 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
      721 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
      781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1561 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2581 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2641 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2701 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2761 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     2821 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     2881 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     2941 cactaaaggg aacaaaagct gggtaccggg ccccccctcg aggtcgacgg tatcgataag
     3001 cttgat
//
