LOCUS       sequence-176197-        3695 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 3695)
  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 3695)
  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: Rab5aa cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     346..368
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     360..377
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     361..377
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     384..403
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        384..402
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      433..442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      437..446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1220..1238)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1221..1238)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1256..1272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1256..1272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1269..1291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1280..1296
                     /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(1304..1310,1311..1328,1329..1334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1349..1370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1487..1504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1658..2246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1738..1757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2417..3208,3209..3277))
                     /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     3040..3059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3278..3382)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3460..3695,1..220))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3597..3618)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
       61 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
      121 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
      181 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc
      241 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      301 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      361 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt
      421 cgcatgctcc cggccgccat ggcggccgcg ggaattcgat taactgtcat ggccaatagg
      481 ggaggagcaa cacgccccaa cgggtccaac gcgggcaata agatctgcca gtttaaactg
      541 gtcttgctag gagagtcagc ggtggggaag tccagcctcg ttctgcgctt cgtcaaaggg
      601 cagtttcacg agtttcagga aagcacaata ggagcggcct ttcttacaca gacactgtgc
      661 ttggatgaca cgacggtaaa gtttgagatc tgggacacag ctggacagga gcgctaccac
      721 agtctggccc ccatgtacta cagaggtgcc caggccgcca ttgtagttta tgacatcacc
      781 aatgaggagt catttgcaag agcaaagaac tgggttaaag agcttcagag gcaggctagc
      841 ccaaacattg tcatcgctct gtctgggaac aaggctgacc ttgccaacaa gagagctgtg
      901 gacttccagg atgctcagtc ttatgcagat gacaacagtt tgctgttcat ggagacgtca
      961 gcaaaaacct ccatgaacgt gaacgagatc ttcatggcca ttgcgaaaaa gttgcctaag
     1021 agtgagcccc aagctgccgg agccaacagc gggcggagta ggggtgtgga cctcacagag
     1081 acagcccaac ccactaaagc cccctgctgc agcaactaac gcaaaacaaa aatccccgta
     1141 atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc ccaacgcgtt
     1201 ggatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat
     1261 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1321 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1381 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1441 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1501 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1561 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1621 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1681 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1741 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1801 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1861 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1921 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1981 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2041 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2101 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2161 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2341 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3001 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa
     3421 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt
     3481 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3541 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3601 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3661 tctatcaggg cgatggccca ctacgtgaac catca
//
