LOCUS       sequence-246543-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High-copy number vector for the co-expression of two genes in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  AUTHORS   Miazzi F, Hoyer C, Sachse S, Knaden M, Wicher D, Hansson BS, 
            Lavista-Llanos S
  TITLE     Optimization of Insect Odorant Receptor Trafficking and Functional 
            Expression Via Transient Transfection in HEK293 Cells.
  JOURNAL   Chem Senses. 2019 Oct 26;44(9):673-682. doi: 10.1093/chemse/bjz062.
  PUBMED    31504297
REFERENCE   2  (bases 1 to 4111)
  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"; doi: "10"; journalName: 
            "Chem Senses"; date: "2019-10-26- 26"; volume: "44"; issue: "9"; 
            pages: "673-682"
COMMENT     Sequence Label: pCMV-BI
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    110..191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        494..797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        798..1001
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     952..972
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1204..1285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1296..1313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1421..1876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1508..1527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1718..1739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1943..1962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2062..2084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2122..2140)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2208..2312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2313..2381,2382..3173)
                     /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(2531..2550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3344..3932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3833..3852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
       61 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgg cagtgaaaaa
      121 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
      181 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
      241 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcctc
      301 tagactgcag cctcaggaga tctgggcccc cgcggcatat gaccggtgaa ttctccaggc
      361 gatctgacgg ttcactaaac gagctctgct tatataggcc tcccaccgta cacgccacct
      421 cgacatactc gagtagttat taatagtaat caattacggg gtcattagtt catagcccat
      481 atatggagtt ccgcgttaca taacttacgg taaatggctc gcctggctga ccgcccaacg
      541 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      601 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      661 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      721 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      781 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      841 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      901 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      961 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     1021 tccgctaggg atcctctagt cagctgacgc gtgctagcgc ggccgcatcg ataagcttgt
     1081 cgacgatatc tccagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     1141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1261 acaaataaag catttttttc actgccccga gcttcctcgc tcactgactc gctgcgctcg
     1321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaga
     1381 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     1441 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1501 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1561 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1621 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1681 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1741 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1801 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1861 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1921 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1981 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2041 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2101 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2161 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2221 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2281 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2401 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2521 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2581 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2701 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3181 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3241 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3301 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3421 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3481 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3541 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3601 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3661 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3721 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3781 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3841 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3901 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3961 tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatcttcaat attggccatt
     4021 agccatatta ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac
     4081 gttgtatcta tatcataata tgtacattta t
//
