LOCUS       sequence-226447-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ARF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5190)
  AUTHORS   Kahn RA, Kern FG, Clark J, Gelmann EP, Rulka C
  TITLE     Human ADP-ribosylation factors. A functionally conserved family of 
            GTP-binding proteins.
  JOURNAL   J Biol Chem. 1991 Feb 5;266(4):2606-14.
  PUBMED    1899243
REFERENCE   2  (bases 1 to 5190)
  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: "J Biol 
            Chem."; date: "1991-02-5"; volume: "266(4)"; pages: "2606-14"
COMMENT     Sequence Label: rk1s
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        47..65
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(685..709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(762..780)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      776..823
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1189..1194,1195..1211,1212..1217))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1244..1262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1330..1434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1435..1503,1504..2295)
                     /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(1653..1672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2466..3054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2955..2974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3208..3225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3240..3379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3305..3324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3465..3487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3481..3672)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5134..5153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
       61 atagggagac cacaacggtt tccctctaga tcacaagttt gtacaaaaaa gcaggcttag
      121 aaggagatag aaccatgggg aacatcttcg ccaacctctt caagggcctt tttggcaaaa
      181 aagaaatgcg catcctcatg gtgggcctgg atgctgcagg gaagaccacg atcctctaca
      241 agcttaagct gggtgagatc gtgaccacca ttcccaccat aggcttcaac gtggaaaccg
      301 tggagtacaa gaacatcagc ttcactgtgt gggacgtggg tggccaggac aagatccggc
      361 ccctgtggcg ccactacttc cagaacacac aaggcctgat cttcgtggtg gacagcaatg
      421 acagagagcg tgtgaacgag gcccgtgagg agctcatgag gatgctggcc gaggacgagc
      481 tccgggatgc tgtcctcctg gtgttcgcca acaagcagga cctccccaac gccatgaatg
      541 cggccgagat cacagacaag ctggggctgc actcactacg ccacaggaac tggtacattc
      601 aggccacctg cgccaccagc ggcgacgggc tctatgaagg actggactgg ctgtccaatc
      661 agctccggaa ccagaagtga taggacccag ctttcttgta caaagtggtg atgatccggc
      721 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      781 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      841 atccggatat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa
      901 gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg
      961 gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga
     1021 tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata accaagccta
     1081 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
     1141 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat
     1201 cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt gatacgccta
     1261 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1381 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1441 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1501 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1561 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1621 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     1681 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1741 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1801 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1861 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1921 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     1981 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2041 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2101 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2161 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2221 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2281 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2341 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2401 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2581 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3061 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3121 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3181 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3241 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     3301 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3361 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3421 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3481 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3541 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     3601 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     3661 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     3721 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     3781 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     3841 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     3901 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     3961 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     4021 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     4081 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     4141 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
     4201 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
     4261 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
     4321 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
     4381 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
     4441 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
     4501 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
     4561 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
     4621 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
     4681 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     4741 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     4801 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     4861 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     4921 aagggcatcg gtcgatcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4981 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5041 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5101 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5161 aaccgcacct gtggcgccgg tgatgccggc
//
