LOCUS       sequence-176726-        3663 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 3663)
  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 3663)
  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: Rab25b cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(127..715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(207..226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(886..1677,1678..1746))
                     /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     1509..1528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1747..1851)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1929..2384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2066..2087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2278..2297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2510..2532
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2524..2541
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2525..2541
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2548..2567
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2548..2566
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2597..2606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2601..2610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3352..3370)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3353..3370)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3388..3404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3388..3404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3401..3423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3412..3428
                     /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(3436..3442,3443..3460,3461..3466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3481..3502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3619..3636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
       61 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1861 ccgaaaagtg ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     1921 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1981 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2041 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2101 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2161 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2221 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2281 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2341 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     2401 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     2461 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     2521 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attgggcccg
     2581 acgtcgcatg ctcccggccg ccatggcggc cgcgggaatt cgatttggaa atatggggtc
     2641 tgatgaggcc tacaacttcg ttttcaaagt ggttcttatt ggagaatccg gtgttggcaa
     2701 aagcaacttg ctgtcacggt tcaccaagaa cgagtttagc cacgacagcc ggacgaccat
     2761 cggcgtggag ttcagcacta gaactgtgca gctgaacgga ctcactatta aagctcagat
     2821 ctgggacacc gctggattgg agcggtaccg cgccatcacc tctgcatatt atcgaggagc
     2881 ggtcggagct ctgcttgttt atgatatttc caagcatctg acgtatgaaa gtgccgagcg
     2941 ctggctgaag gagctctacg atcacgcaga tccacacatc gtagtgatgc tggtgggaaa
     3001 taaatcagac ctggcagccg tgcgctccgt tcctacagag gacgccaaag actttgcaga
     3061 gaagaatggt cttctgttca tggaaacctc agctctggag tcagtaaatg tggaagcagc
     3121 gttcaacact gtgcttacag aaatccacaa gaaggtgaac agcaaagaag tgacccgcgg
     3181 ctccatcagt gcagtcacgt tgtctcaacc taaaactcca gcagaaaccc aggaggagaa
     3241 gaaaacctgc tgtaaaaact tgtgaaagag taatcactag tgaattcgcg gccgcctgca
     3301 ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta ttctatagtg
     3361 tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3601 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3661 agc
//
