LOCUS       sequence-168000-        3689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used for in situ hybridisation or other purposes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3689)
  AUTHORS   Hall TE, Martel N, Lo HP, Xiong Z, Parton RG
  TITLE     A plasmid library of full-length zebrafish rab proteins for in vivo 
            cell biology.
  JOURNAL   Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 
            10.1080/21592799.2017.1301151. eCollection 2017.
  PUBMED    28396820
REFERENCE   2  (bases 1 to 3689)
  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: "Cell 
            Logist."; date: "2017-03-1"; volume: "7(1):e1301151. doi"; pages: " 
            10.1080/21592799.2017.1301151. eCollection 2017"
COMMENT     Sequence Label: Rab4a cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        210..314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(315..383,384..1175)
                     /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(533..552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1346..1934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1835..1854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2088..2105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2222..2243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2258..2263,2264..2281,2282..2288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2296..2312
                     /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     2301..2323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2320..2336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2320..2336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2354..2372
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2354..2371
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3140..3149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3144..3153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3183..3202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3184..3202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3209..3226)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3209..3225)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3218..3240)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(3366..3689,1..132)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3453..3472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3663..3684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
       61 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      121 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      181 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      241 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      301 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      361 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      421 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      481 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      541 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      601 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      661 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      721 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      781 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      841 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      901 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      961 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1021 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1081 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1141 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1201 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1261 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1981 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2041 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2101 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2161 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2221 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     2281 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     2341 attacgccaa gctatttagg tgacactata gaatactcaa gctatgcatc caacgcgttg
     2401 ggagctctcc catatggtcg acctgcaggc ggccgcgaat tcactagtga ttagcgccag
     2461 tcatgtcaga gacatacgat ttcttgttta agtttcttgt gattgggaat gctggaacag
     2521 gaaaatcctg tcttcttcat cagttcatag aaaaaagatt caaagatgac tcaaatcaca
     2581 ctattggggt tgaatttggt tcgaagataa tcagcgtggt caataagttt gtaaagctcc
     2641 agatctggga cacagcagga caggagcgtt tcaggtctgt aactaggagt tattacagag
     2701 gcgcagcagg ggctcttcta gtctatgaca tcactagtcg agaaacatac aacgctctaa
     2761 ccaactggct gacagatgcc cggatgttgg ccagtcagaa catagtgatc atactatgtg
     2821 gaaacaagaa ggatctggat gctgaccgcg aggtgacctt tctggaggct tcacgcttcg
     2881 ctcaggagaa tgagcttatg tttttggaga ccagtgctct gacaggggag aatgtggagg
     2941 aagctttcgt ccagtgtgct cgaaaaatcc ttaacaaaat tgaatcagga gagctggacc
     3001 cagagcggat gggttcagga atccagtatg gagacgcggc tcttcgacag ctgcgttcac
     3061 ccagacgggc acaagccgag agcatccagg agtgtggctg ttaacacacg tccctctgaa
     3121 cagaaatcga attcccgcgg ccgccatggc ggccgggagc atgcgacgtc gggcccaatt
     3181 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3361 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3421 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3481 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3541 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3601 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3661 tagtggactc ttgttccaaa ctggaacaa
//
