LOCUS       sequence-175305-        3657 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 3657)
  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 3657)
  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: Rab7a cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        651..669
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     651..668
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1405..1414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1409..1418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1448..1467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1449..1467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1474..1491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1474..1490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1483..1505)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1631..2086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1718..1737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2164..2268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2269..2337,2338..3129)
                     /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(2487..2506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3300..3657,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct atttaggtga
      661 cactatagaa tactcaagct atgcatccaa cgcgttggga gctctcccat atggtcgacc
      721 tgcaggcggc cgcgaattca ctagtgattg gtcctcagca gctacaggtc tctgctgatg
      781 gtttggctct atcgttccgg tctagtttaa ttggctcggg gaactcgttg tataactcca
      841 cttcagtttc ctgtttgagt gcattgcgcg cgatggtctg gaatgcctgc tctacgttga
      901 tggcctcctt tgcactggtc tcaaaatatg ggatgttgtt cttactctga caccaagctt
      961 gtgctcgctt agttgttacc tgtctattct ccagatcgat tttgttgccc agcaccacga
     1021 aagggaagtt ttctgggtct cgaggactgg cctggatcag aaactcatcc ctccagctgt
     1081 ccagtgtctt gaaggtgttc ggggcagtga cgtcaaacac caggacgcag cagtcagcac
     1141 cacgatagaa agctacaccc agagactgaa accgctcctg acctgctgtg ccccatatct
     1201 gcattgtgac aagccggtcg tcaaccatca cttcttttgt caggaaatct gctcctatag
     1261 tggctttata ctggttactg aactttttat tcacatactg gttcattaga gatgtttttc
     1321 caaccccgga atctccaaga atgatcacct tcagaagaac tttcttcctt gatgtcatag
     1381 tgtcagagca atcgaattcc cgcggccgcc atggcggccg ggagcatgcg acgtcgggcc
     1441 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     1501 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1561 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1621 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1681 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1741 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1801 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1861 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1921 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1981 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2041 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2101 ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat
     2161 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2221 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2281 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3121 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3301 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3361 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3421 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3481 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3541 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3601 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacg
//
