LOCUS       sequence-176657-        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: Rab5b cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..48)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     239..258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     471..493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     485..502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     486..502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     509..528
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        509..527
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      562..571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1334..1352)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1335..1352)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1370..1386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1370..1386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1383..1405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1394..1410
                     /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)."
     promoter        complement(join(1418..1424,1425..1442,1443..1448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1463..1484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1601..1618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1772..2360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1852..1871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2531..3322,3323..3391))
                     /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     3154..3173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3392..3496)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3574..3684,1..345))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
       61 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
      361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      481 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattgggccc
      541 gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat tcgattggcc acccttccta
      601 catcatgagt acccgtggga caggcagggc caatgggaac ctaccgcaga ccaagatctg
      661 ccagtttaaa ctggtgctgc tgggagacat ggccgtcggc aagtccagcc tggtgctgcg
      721 ctttgtgaag ggacagtttg atgagtttca ggagaccacc attggagctg cattcttggc
      781 ccagtctgtt tgtctggacg acactacagt gaagtttgag atttgggaca cagccggaca
      841 agagcgctac cacagcttgg ctcccatgta ctaccgtgga gcccaggcag ccattgtagt
      901 gtttgatatc actaaacctg aaacatttga gcgagcaaaa gcatgggtga aagagctgca
      961 gcggcaggcc agtcccaaca tagtcattgc tcttgcagga aacaaagccg atctagcaga
     1021 caagagactt gtggagtatg aggaagccca gacatatgca gaagacacag gtttactgtt
     1081 catggaaacc tctgccaaga ctgcaatgaa tgtaaacgag ttgttcctgg ccattgcaaa
     1141 aaagatgcca aaaacagaca cccaaaaccc tacgcatgct gcgcgacaca ggggcgtgaa
     1201 cctgcaggac ccagacgcac agtccactcg aagctgctgt gggaactaaa ggcaatcact
     1261 agtgaattcg cggccgcctg caggtcgacc atatgggaga gctcccaacg cgttggatgc
     1321 atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg tcatagctgt
     1381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2281 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2341 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2401 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2461 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2521 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2581 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2641 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2701 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2761 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2821 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2881 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2941 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3001 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3061 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3121 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3181 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3241 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3301 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3361 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3421 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3481 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg
     3541 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     3601 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3661 aaatccctta taaatcaaaa gaat
//
