LOCUS       sequence-8145-ex        5206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5206)
  AUTHORS   Fiordalisi JJ, Johnson RL 2nd, Ulku AS, Der CJ, Cox AD
  TITLE     Mammalian expression vectors for Ras family proteins: generation and
            use of expression constructs to analyze Ras family function.
  JOURNAL   Methods Enzymol. 2001;332:3-36. doi: 10.1016/s0076-6879(01)32189-4.
  PUBMED    11305105
REFERENCE   2  (bases 1 to 5206)
  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: "Methods 
            Enzymol."; date: "2001"; volume: "332:3-36. doi"; pages: " 
            10.1016/s0076-6879(01)32189-4"
COMMENT     Sequence Label: pBABE HAII
FEATURES             Location/Qualifiers
     source          1..5206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1373..1399
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1379..1405
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     primer_bind     complement(1445..1465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1450..1779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1630..1765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1789..2388
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1789..1808)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2285..2305
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2487..3080
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3281..3298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3452..4040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3532..3551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4218..5009,5010..5078))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4841..4860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5079..5183)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatca atggcttcaa gttatccata
     1381 tgatgtgccc gactatgcct cacttggagg accctctgga tccggaattc tccgtcgacc
     1441 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1501 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1561 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1621 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1681 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     1741 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac
     1801 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     1861 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     1921 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     1981 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     2041 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     2101 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     2161 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     2221 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     2281 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgagtgc
     2341 ccgaaggacc gcgcgacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     2401 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgataaa ataaaagatt
     2461 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     2521 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     2581 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     2641 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     2701 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     2761 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     2821 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     2881 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     2941 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     3001 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     3061 gtcagcgggg gtctttcatt tccgacttgt ggtctcgctg ccttgggagg gtctcctctg
     3121 agtgattgac tacccgtcag cgggggtctt cacatgcagc atgtatcaaa attaatttgg
     3181 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     3241 gcggggagag gcggtttgcg tattggcgct cttccgcttc ctcgctcact gactcgctgc
     3301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4141 agatcctttt aaattaaaaa tgaagtttgc ggccgcaaat caatctaaag tatatatgag
     4201 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4261 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4321 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4381 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4441 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4501 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4561 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4621 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4681 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4741 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4801 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     4861 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4921 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4981 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5041 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5101 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5161 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
