LOCUS       sequence-176581-        3684 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 3684)
  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 3684)
  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: Rab4b cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    243..264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(279..284,285..302,303..309)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    317..333
                     /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     322..344
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     341..357
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     341..357
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        375..393
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     375..392
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1156..1165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1160..1169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1199..1218)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1200..1218)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1225..1242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1225..1241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1234..1256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1382..1837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1469..1488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1679..1700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1915..2019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2020..2088,2089..2880)
                     /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(2238..2257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3051..3639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3540..3559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      121 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
      181 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      241 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      301 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      361 gattacgcca agctatttag gtgacactat agaatactca agctatgcat ccaacgcgtt
      421 gggagctctc ccatatggtc gacctgcagg cggccgcgaa ttcactagtg attgggaccg
      481 acatgtcaga gacgtacgat ttcctgttca agttcctggt gattggcagc gccgggactg
      541 ggaaatcatg cctccttcac cagttcatag agaacaagtt caaacaggac tccaatcaca
      601 ccatcggcgt tgagttcggc tccagggttg tcaatgttgg tggcaaaaca gtcaaactgc
      661 agatctggga cacggctgga caggagcgct ttaggtcagt gacccgcagt tactatcgtg
      721 gagcagcagg agctcttctc gtgtatgaca tcacaagtcg ggagacctac aatgctctga
      781 ctaactggtt gacagacgca aggacactgg ccagtcccaa catcgtcatc atcctgtgcg
      841 gaaataagaa ggatctgggc gcagaccgtg aagtcacttt cctagaggca tctcgcttcg
      901 cccaggaaaa tgagctgatg tttttagaga cgagtgcttt gactggggag aatgtcgagg
      961 aagcctttct taaatgcgct cgcagcatcc tcaacaagat tgaatcaggt gagctggacc
     1021 cagagcggat gggttctggg attcagtatg gcgacgcgtc cctgcggcag atccgacagc
     1081 cccggggctc tgctgcacag accaagcagc agtgtaactg ttagggctct gaagaccctt
     1141 aatcgaattc ccgcggccgc catggcggcc gggagcatgc gacgtcgggc ccaattcgcc
     1201 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1261 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1321 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1381 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1441 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1501 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1561 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1621 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1681 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1741 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1801 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     1861 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     1921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1981 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2041 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2101 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2161 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2221 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2281 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2341 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2401 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2461 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2521 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2581 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2641 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2701 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2761 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2821 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2881 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2941 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3181 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3661 ttacggttcc tggccttttg ctgg
//
