LOCUS       sequence-10188-e        7147 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 7147)
  AUTHORS   Juven-Gershon T, Cheng S, Kadonaga JT
  TITLE     Rational design of a super core promoter that enhances gene 
            expression.
  JOURNAL   Nat Methods. 2006 Nov . 3(11):917-22.
  PUBMED    17124735
REFERENCE   2  (bases 1 to 7147)
  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: "Nat 
            Methods. 2006 Nov . 3(11):917-22."
COMMENT     Sequence Label: SCP2-CMVenh-luc
FEATURES             Location/Qualifiers
     source          1..7147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      29..38
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             35..1687
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(101..119)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(1704..1722)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1705..1722)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1742..1759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1748..1972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2018..2531
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2105..2124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2315..2336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2554..2574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2559..2889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2740..2875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2956..3750
                     /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(3010..3029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3620..3639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3924..4058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3961..3980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4015..4034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4090..4106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4090..4106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4103..4125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4114..4130
                     /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(4138..4144,4145..4162,4163..4168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4183..4204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4321..4338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4492..5080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4572..4591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5251..6042,6043..6111))
                     /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     5874..5893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6112..6216)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6291..6310)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6482..6861
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6862..7065
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7016..7036
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7062..7086
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     7127..7146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7127..7145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
       61 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
      121 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
      181 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
      241 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
      301 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
      361 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
      421 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
      481 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
      541 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
      601 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
      661 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
      721 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
      781 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
      841 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
      901 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
      961 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     1021 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     1081 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     1141 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     1201 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     1261 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     1321 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     1381 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     1441 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     1501 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     1561 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     1621 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     1681 cgtgtaattc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     1741 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1801 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1861 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1921 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1981 gcggaaagaa ccagctgggg ctcgaggggg gatccccacg cgccctgtag cggcgcatta
     2041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2161 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2221 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2281 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2341 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     2401 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2461 acgtttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta
     2521 tcaaccgggg tgggtaccga gctcgaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2581 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2881 ttttgcaaaa agctcccggg agcttggata tccattttcg gatctgatca agagacagga
     2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3121 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3481 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3901 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3961 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4021 tgtccaaact catcaatgta tcttatcatg tctggatccc gtcgacctcg agagcttggc
     4081 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4141 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4681 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4921 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6241 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     6301 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6361 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6421 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6481 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6541 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6601 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6661 actttccatt gacgtcaatg ggtggactat ttacggtaaa ctgcccactt ggcagtacat
     6721 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6781 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6841 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     6901 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     6961 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     7021 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
     7081 cagatcgcct ggagacgtcg agccgagtgg ttgtgcctcc atagaataat acgactcact
     7141 ataggga
//
