LOCUS       sequence-176248-        3703 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 3703)
  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 3703)
  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: Rab5ab cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    114..135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(150..155,156..173,174..180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..204
                     /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     193..215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     212..228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     212..228
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        246..264
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     246..263
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1046..1055
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1050..1059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1089..1108)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1090..1108)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1115..1132)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1115..1131)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1124..1146)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1272..1727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1359..1378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1569..1590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1805..1909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1910..1978,1979..2770)
                     /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(2128..2147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2941..3529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3430..3449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3683..3700
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
       61 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      121 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      181 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      241 aagctattta ggtgacacta tagaatactc aagctatgca tccaacgcgt tgggagctct
      301 cccatatggt cgacctgcag gcggccgcga attcactagt gattgaggtg aggagcgtgg
      361 tttaggtgct acagcagggg cctttggcgg gctgtgctgt ttctgttaga tccactcctc
      421 tgttgcgggc tgtgttggca cctgcgggct ggggctcgtt cttcgggagc ttctttgcga
      481 tcgccataaa aatctcactg acgttcattg aagtctttgc cgacgtttcc ataaagagca
      541 aactgttgtc atctgcatat gattgcgcat cctgaaagtc caacgctctc ttgttggcaa
      601 ggtctgcctt gttcccagcc aaagctatta caatatttgg actggcttgt ctttgaagct
      661 ctttcaccca gtttttcgca cgtgcaaatg attcttcatt tgtgatgtca tacaccacta
      721 tggccgcctg tgctcctctg taatacattg gggccagact gtggtagcgc tcctggccag
      781 ctgtatccca gatctcaaac ttcgccgttg tgtcatccag acataatgtt tgagtcaaga
      841 aggcggctcc tattgtgctt tcctgaaact catgaaactg tccttttaca aagcgcagca
      901 ctagactcga cttccccaca gcagactctc ccagcagcac taatttgaac tgacagattt
      961 tgttgccggt gttcggcccg ttaggtctcg ttgctccacc tcttcctgcc atgacccaaa
     1021 accccaatct aatcgaattc ccgcggccgc catggcggcc gggagcatgc gacgtcgggc
     1081 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     1141 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1201 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1261 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1621 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1681 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     1741 tttctcctta cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa
     1801 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1861 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1921 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1981 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2041 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2101 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2161 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2221 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2281 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2341 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2401 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2461 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2521 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2581 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2641 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2701 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2761 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2821 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2881 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2941 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3001 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3061 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3121 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3181 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3241 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3301 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3361 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3421 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3481 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3541 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3601 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3661 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaa
//
