LOCUS       sequence-55746-e        6853 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 6853)
  AUTHORS   Yang T, Puckerin A, Colecraft HM
  TITLE     Distinct RGK GTPases differentially use alpha1- and auxiliary 
            beta-binding-dependent mechanisms to inhibit CaV1.2/CaV2.2 channels.
  JOURNAL   PLoS One. 2012;7(5):e37079. Epub 2012 May 10.
  PUBMED    22590648
REFERENCE   2  (bases 1 to 6853)
  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: "PLoS One.";
            date: "2012"; volume: "7(5)"; pages: "e37079. Epub 2012 May 10"
COMMENT     Sequence Label: CFP_Rem_pcDNA4_HisMaxC
FEATURES             Location/Qualifiers
     source          1..6853
                     /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"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    917..1079
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1092..1109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1113..1145
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1149..1157,1158..1172)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1179..1181,1182..1892)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1221..1242)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1482..1501)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1829..1850
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2860..2877)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2866..3090
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3136..3564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3433..3454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3573..3593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3578..3908
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3759..3894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3821..3840
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3956..4003
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4022..4396
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4526..4647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4563..4582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4617..4636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4696..4712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4696..4712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4709..4731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4720..4736
                     /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(4744..4750,4751..4768,4769..4774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4789..4810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4927..4944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5098..5686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5178..5197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5857..6648,6649..6717))
                     /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     6480..6499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6718..6822)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttagcg cagaggcttg gggcagccga gcggcagcca ggccccggcc
      961 cgggcctcgg ttccagaagg gagaggagcc cgccaaggcg cgcaagagag cgggctgcct
     1021 cgcagtccga gccggagagg gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca
     1081 tggggggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     1141 tgggtcggga tctgtacgac gatgacgata aggtaccagt gagcaagggc gaggagctgt
     1201 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1261 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1321 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctggggcg
     1381 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1441 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1501 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1561 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacatcagcc
     1621 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1681 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1741 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     1801 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1861 ggatcactct cggcatggac gagctgtaca agggatccac tcttaacacc cagcaggaag
     1921 caaagaccac tctgcgacgt cgagcaagca caccactacc cctgtcgtcc aggggccacc
     1981 agcctggccg cctgtgcaca gcaccctctg ctccatccca gcatccccga ctgggtcagt
     2041 cagtctctct caaccctccc gtccggaaac cctcccctgc ccaggatggg tggtcctctg
     2101 aatccagcga ctccgaaggc tcttgggagg cactctaccg ggtggtgctt cttggagacc
     2161 ctggtgttgg gaagactagc ctggctagcc tcttcgcaga gaaacaagac cgagaccctc
     2221 acgagcagct gggaggtgtg tacgagagaa cgctgtcagt ggacggagag gacaccactc
     2281 tggtggtcat ggacacctgg gaggctgaga aactggatga aagctggtgc caggagtcat
     2341 gcctgcaggc aggcagcgcc tacgtcatcg tgtactccat agcggatcgc agcagctttg
     2401 agagcgcctc ggagctccga atccagctga ggcgcacgca tcaggccaac cacgtgccca
     2461 tcatcctggt gggcaacaag gccgacctgg cccgctgccg ggaagtctcg gtagaagagg
     2521 gtcgcgcctg cgccgtggtg ttcgactgca agttcatcga gacttcggcc actctgcagc
     2581 acaatgtgac agagctcttc gagggcgtgg tgcgccaact gcgcctgcgc cgccaggaca
     2641 atgcagcccc ggagacacct tcacctcgac gacgggccag cttgggccag cgcgcccgcc
     2701 gattcctggc acgtctgaca gcacgcagcg cacgacgccg ggcacttaaa gcccgctcca
     2761 agtcctgcca caacctggct gtgctctgag aattctgcag atatccagca cagtggcggc
     2821 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2881 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2941 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3001 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3061 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3121 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     3301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3481 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     3541 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3601 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3661 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3781 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3841 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3901 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3961 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     4021 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     4081 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     4141 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     4201 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     4261 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     4321 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     4381 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4441 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4501 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4561 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4621 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4681 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4741 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4801 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4861 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4921 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4981 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5101 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5161 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5221 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5281 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5341 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5401 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5461 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5521 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5581 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5641 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5701 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5761 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5821 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5881 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5941 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6001 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6061 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6121 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6181 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6241 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6301 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6361 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6421 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6481 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6541 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6601 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6661 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6721 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6781 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6841 gccacctgac gtc
//
