LOCUS       sequence-176447-        3679 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 3679)
  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 3679)
  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: Rab32a cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     366..383
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(536..541,542..559,560..566)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    574..590
                     /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     579..601
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     598..614
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     598..614
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        632..650
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     632..649
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1408..1417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1412..1421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1451..1470)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1452..1470)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1477..1494)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1477..1493)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1486..1508)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1634..2089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1721..1740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1931..1952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2167..2271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2272..2340,2341..3132)
                     /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(2490..2509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3303..3679,1..212)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
       61 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      601 gaaacagcta tgaccatgat tacgccaagc tatttaggtg acactataga atactcaagc
      661 tatgcatcca acgcgttggg agctctccca tatggtcgac ctgcaggcgg ccgcgaattc
      721 actagtgatt tgacattttg gcagcatggc aggcgggtcc gtgtcggagt gtaaggagta
      781 tctgtttaaa gttttggtta tcggggaact tggtgtggga aagactagca tcatcaagcg
      841 atatgttcac cagctcttct cgcaacacta cagagcgacc ataggcgtgg attttgctct
      901 caaagtctta aactgggaca gcaaaacact agtacgacta cagctgtggg acatcgcagg
      961 tcaggagcgc tttggaaaca tgaccagagt ttactacaaa gaggcagtgg gtgcatttat
     1021 tgtgtttgac attactcgag gctccacgtt tgaggctgtt tcaaaatgga agcatgattt
     1081 ggacaccaag gtgaaactgg ccaatggcaa tcctatacct tcagtccttc tggccaataa
     1141 atgcgaccag aagaaagacg gatccaacaa ttcctcactc atggacaact tctgcaagga
     1201 ggctggcttt ttgggttggt ttgaaacatc agccaaggag aacatcaacg tggacgaggc
     1261 tgcccggttc ttggtggaga acatccttct caatgataaa ggtcttccgt acgaagagaa
     1321 caatggtgac aagattaaac tagaccaaca aactgctcct gctgagagca agtcaggttg
     1381 ctgctagtgc ggaatcgaat tcccgcggcc gccatggcgg ccgggagcat gcgacgtcgg
     1441 gcccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     1501 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1561 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1621 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1681 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1741 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1801 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1861 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1921 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1981 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2041 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     2101 attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga
     2161 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2221 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2281 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2341 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2401 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2461 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2521 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2581 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2641 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2701 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2761 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2821 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2881 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2941 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3001 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3061 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3421 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3661 acctacaccg aactgagat
//
