LOCUS       sequence-200395-        4973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A modified pcDNA3.1- vector with reduced expression activity of a 
            truncated CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4973)
  AUTHORS   Kainrath S, Stadler M, Reichhart E, Distel M, Janovjak H
  TITLE     Green-Light-Induced Inactivation of Receptor Signaling Using 
            Cobalamin-Binding Domains.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Apr 10;56(16):4608-4611. doi: 
            10.1002/anie.201611998. Epub 2017 Mar 20.
  PUBMED    28319307
REFERENCE   2  (bases 1 to 4973)
  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: 
            "10.1002/anie.201611998"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2017-04-10- 10"; volume: "56"; issue: "16"; pages: 
            "4608-4611"
COMMENT     Sequence Label: pcDNA3.1(-)_CMVtrunc_877
FEATURES             Location/Qualifiers
     source          1..4973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     409..428
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        409..427
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(567..584)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    573..797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      843..1271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(930..949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1140..1161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1280..1300)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1285..1614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1465..1600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1527..1546
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1681..2475
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1735..1754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2345..2364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2649..2770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2686..2705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2740..2759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2819..2835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2819..2835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2832..2854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2843..2859
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2867..2873,2874..2891,2892..2897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2912..2933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3050..3067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3221..3806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3301..3320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3977..4768,4769..4837))
                     /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     4600..4619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4838..4942)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tgggaggtct atataagcag
      361 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      421 ctatagggag acccaagctg gctagcgttt aaacgggccc tctagactcg agcggccgcc
      481 actgtgctgg atatctgcag aattccacca cactggacta gtggatccga gctcggtacc
      541 aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
      601 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
      661 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
      721 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
      781 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
      841 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      901 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      961 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1021 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1081 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1141 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1201 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1261 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1321 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1381 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1441 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1501 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     1561 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     1621 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     1681 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1741 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1801 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1861 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     1921 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     1981 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2041 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2101 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2161 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     2221 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2281 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2341 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2401 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2461 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     2521 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     2581 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     2641 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2761 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     2821 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2881 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2941 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3001 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3661 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3721 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     3781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3901 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3961 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4141 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4501 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4801 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4861 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4921 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
