LOCUS       sequence-403025-        5219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARL14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5219)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5219)
  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"
COMMENT     Sequence Label: pET3-ARL14-His6
FEATURES             Location/Qualifiers
     source          1..5219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             747..764
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(818..836)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      832..879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1245..1250,1251..1267,1268..1273))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1300..1318)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1386..1490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1491..1559,1560..2351)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1709..1728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2522..3110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3011..3030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3264..3281
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3296..3438
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3364..3383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3524..3546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3540..3731)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5189..5208
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
       61 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      121 attttgttta actttaagaa ggagatatac atatgggttc gctgggttct aaaaatccgc
      181 aaaccaaaca agcccaagtt cttcttttgg gacttgactc agctgggaag tctactctcc
      241 tttataaatt aaagcttgct aaggatatta ccaccatccc tacaataggt ttcaatgtgg
      301 aaatgatcga gttggaaagg aatctttcac tcacagtctg ggatgttgga ggacaggaaa
      361 aaatgagaac tgtttggggc tgttactgtg agaacaccga tgggctggtg tatgttgtgg
      421 acagtacaga caaacagcga ctggaagagt ctcagagaca gtttgagcac attttgaaga
      481 atgaacacat taaaaatgtg cctgttgttc tattagccaa caaacaagac atgcctggag
      541 ctctgactgc tgaggacatc accagaatgt tcaaagtgaa gaagctttgc agtgaccgga
      601 actggtatgt gcaaccctgc tgtgccctca caggggaggg gctggcccag gggttcagga
      661 aattaactgg atttgtgaag agccacatga aatcaagagg agacactttg gcgttcttca
      721 agcagaacgg cagcggcagc ggcagccatc accatcacca tcactaagga tccggctgct
      781 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      841 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      901 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
      961 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     1021 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     1081 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     1141 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     1201 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     1261 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     1321 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1501 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1561 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1621 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1681 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1741 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1801 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1861 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1921 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1981 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2041 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2101 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2161 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2221 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2281 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2341 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2401 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2461 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2641 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3181 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3241 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3301 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatat ggtgcactct
     3361 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3421 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3481 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3541 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3601 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3661 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3721 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3781 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3841 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     3901 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     3961 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4021 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4081 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     4141 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     4201 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     4261 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     4321 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     4381 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     4441 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     4501 tcagcggtcc agtgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     4561 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     4621 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     4681 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     4741 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     4801 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     4861 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4921 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4981 agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     5041 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     5101 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5161 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaacc
//
