LOCUS       sequence-176225-        3707 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 3707)
  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 3707)
  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: Rab5c cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(885..1676,1677..1745))
                     /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     1508..1527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1746..1850)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1928..2383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2065..2086)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2277..2296
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2509..2531
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2523..2540
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2524..2540
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2547..2566
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2547..2565
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2596..2605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2600..2609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3395..3413)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3396..3413)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3431..3447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3431..3447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3444..3466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3455..3471
                     /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(3479..3485,3486..3503,3504..3509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3662..3679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1861 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     1921 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1981 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2041 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2101 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2161 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2221 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2281 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2341 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     2401 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2461 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2521 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga
     2581 cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gattctcctg ttgtccagcc
     2641 tgttggcatg gcggggcgag gtggaccagc gcggaccaac ggtgccgccc ccgtcgggaa
     2701 caaaatctgc cagtttaagc ttgtgttgct gggagagtct gcagtaggca agtccagcct
     2761 ggtgctgcgc ttcgtcaaag gccagtttca cgagtaccag gagagcacca ttggagctgc
     2821 tttcctcaca cagacactct gcttggacga tacgacagta aagtttgaga tctgggacac
     2881 ggccggacag gagcggtatc acagtttggc ccctatgtat tacagaggag cccaggcagc
     2941 catcgtcgtc tacgacatta ctaacacaga cacatttact agagcaaaga actgggtgaa
     3001 ggagctgcag cgccaggcca gtccaaatat cgttattgct ctggcaggca acaaagcaga
     3061 tctggccaac aagagagccg ttgactttca ggaggcccag gcatacgcag atgacaacag
     3121 cctactcttc atggagacgt ctgcaaagac cgccatgaac gtcaatgaga tcttcatggc
     3181 tatcgccaag aagcttccga aaagcgagcc gcagggtgga gcgggcagcg ggggacgggc
     3241 ccgaggaggg gtggacttac aggagacggc gcctcaggga agaagcggcc agtgctgtgg
     3301 aggcggaaac taacaatcac tagtgaattc gcggccgcct gcaggtcgac catatgggag
     3361 agctcccaac gcgttggatg catagcttga gtattctata gtgtcaccta aatagcttgg
     3421 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3481 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3601 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcg
//
