LOCUS       sequence-226644-        5187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ARF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  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 5187)
  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: rk3s
FEATURES             Location/Qualifiers
     source          1..5187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     105..124
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        105..123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    151..175
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(740..764)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(817..835)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      831..878
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1244..1249,1250..1266,1267..1272))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1299..1317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1385..1489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1490..1558,1559..2350)
                     /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(1708..1727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2521..3109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3010..3029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3263..3280
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3295..3434
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3360..3379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3520..3542
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3536..3727)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
       61 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
      121 agggagacca caacggtttc cctctagatc acaagtttgt acaaaaaagc aggcttagaa
      181 ggagatagaa ccatgggcct cactatctcc tccctcttct cccgactatt tggcaagaag
      241 cagatgcgca ttttgatggt tggattggat gctgctggca agacaaccat tctgtataaa
      301 ctgaagttag gggagatagt caccaccatt cctaccattg gttttaatgt ggaaacagta
      361 gaatataaga acatttgttt cacagtatgg gatgttggtg gtcaagatag aattaggcct
      421 ctctggaagc attacttcca gaatacccag ggtcttattt ttgtggtaga tagcaacgat
      481 cgtgaaagaa ttcaggaagt agcagatgag ctgcagaaaa tgcttctggt agatgaattg
      541 agagatgcag tgctgctact ttttgcaaac aaacaggatt tgccaaatgc tatggccatc
      601 agtgaaatga cagataaact agggcttcag tctcttcgta acagaacatg gtatgttcaa
      661 gccacttgtg caacacaagg aactggtctg tatgaaggac ttgactggct gtcaaatgag
      721 ctttcaaaac gttaatagga cccagctttc ttgtacaaag tggtgatgat ccggctgcta
      781 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      841 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
      901 gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
      961 gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg
     1021 catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg
     1081 tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
     1141 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
     1201 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
     1261 ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt
     1321 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1381 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1441 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1501 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1561 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1621 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1681 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1741 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1801 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1861 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1921 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1981 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2041 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2101 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2161 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2221 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2281 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2341 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2641 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3181 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3241 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3301 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3361 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3421 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3541 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3601 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3661 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3721 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3781 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3841 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3901 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3961 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4021 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4081 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4141 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     4201 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     4261 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     4321 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     4381 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     4441 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     4501 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     4561 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     4621 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     4681 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4741 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4801 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4861 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4921 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4981 catcggtcga tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     5041 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     5101 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     5161 cgaagtggcg agcccgatct tccccat
//
