LOCUS       sequence-176732-        3658 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 3658)
  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 3658)
  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: Rab13 cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(163..751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(243..262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(922..1713,1714..1782))
                     /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     1545..1564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1783..1887)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1965..2420)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2102..2123)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2314..2333
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2546..2568
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2560..2577
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2561..2577
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2584..2603
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2584..2602
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2633..2642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2637..2646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3383..3401)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3384..3401)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3419..3435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3419..3435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3432..3454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3443..3459
                     /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(3467..3473,3474..3491,3492..3497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3512..3533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(3650..3658,1..9))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
       61 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1201 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
     1921 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     1981 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2041 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2101 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2161 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2221 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2281 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2341 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2401 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2461 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2521 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2581 ttgtaatacg actcactata gggcgaattg ggcccgacgt cgcatgctcc cggccgccat
     2641 ggcggccgcg ggaattcgat tacccagtcg aaacgaacaa tggcaaagaa gtacgacttc
     2701 ctctttaaat tattgcttat cggcgatagc ggtgttggca agacatgttt gatcatccgt
     2761 ttcgcagagg acaattttaa ttcaacgtac atctcaacca tcggtattga cttcaaagtg
     2821 aagaccattg aggtggaagg aaagaaagtg aagctgcaag tctgggacac agcaggacag
     2881 gagaggttta agaccatcac cactgcatac tacagagggg ccatgggaat cattctcgtg
     2941 tatgacataa ctgatgaaaa atcctatgag aatattcaga actggatgaa gagcatcaaa
     3001 gaaaatgcat cagcgggtgt aagtcgaatg ttactgggta acaagtgtga cattgaggcc
     3061 aagaggaaag tctcaaaaga gactggtgag aagctcgcaa aggaacatgg tattcgattc
     3121 tttgagacta gtgccaaatc gagcatcaac gttgaggagt cttttacttc tcttgctcga
     3181 gacattttgc tgaagtcaaa taagaagccg ggtccatctg gacgtgaggt caaacttacc
     3241 agcacagaga aaaagtcctc ctccaaatgt cttcttcttt aggcaaacca ttttccagag
     3301 gaaatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc
     3361 gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt
     3421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcac
//
