LOCUS       sequence-257009-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EcSSPB-micro (Escherichia coli) preceded by restriction 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5706)
  AUTHORS   Tichy AM, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H
  TITLE     Engineering Strategy and Vector Library for the Rapid Generation of 
            Modular Light-Controlled Protein-Protein Interactions.
  JOURNAL   J Mol Biol. 2019 May 29. pii: S0022-2836(19)30317-1. doi: 
            10.1016/j.jmb.2019.05.033.
  PUBMED    31150735
REFERENCE   2  (bases 1 to 5706)
  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: "J Mol Biol.
            2019 May 29. pii: S0022-2836(19)30317-1. doi: 
            10.1016/j.jmb.2019.05.033."
COMMENT     Sequence Label: pOVC1_X_EcSSPB-nano
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        132..511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     666..686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     760..779
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        760..778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             834..1163
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     primer_bind     complement(1194..1211)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1200..1424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1470..1898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1557..1576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1767..1788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1907..1927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1912..2241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2092..2227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2154..2173
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2308..3102
                     /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(2362..2381)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2972..2991
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3276..3397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3313..3332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3367..3386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3446..3462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3446..3462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3459..3481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3470..3486
                     /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(3494..3500,3501..3518,3519..3524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3539..3560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3677..3694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3928..3947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4607..5398,5399..5467))
                     /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     5230..5249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5468..5572)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5647..5666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
       61 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      121 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      481 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      541 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      601 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      661 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      721 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
      781 gacccaagct gaccatggcc accggtgaat tctcaggctc tggatccccc gggagctccc
      841 cgaaacgccc taagctgctg cgtgaatatt acgattggct ggttgataac agctttaccc
      901 catatctggt ggtggatgcc acatacctgg gcgtgaacgt gcccgtggag tatgtgaaag
      961 acggtcagat cgtgctgaat ctgtctgcaa gtgcgaccgg caacctgcaa ctgacaaatg
     1021 attttatcca gttcaacgcc cgctttaagg gcgtgtctcg tgaactgtat atcccgatgg
     1081 gtgccgctct ggccatttac gctcgcgaga acggcgatgg tgtgatgttc gaaccagaag
     1141 aaatctatga cgagctgaat attggtcccg ggtaagttta aaccgctgat cagcctcgac
     1201 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1261 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1321 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1381 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1441 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     1501 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1561 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1621 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1681 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1741 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1801 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1861 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     1921 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1981 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2041 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2101 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2161 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2221 ggaggcctag gcttttgcaa aaagctctcg ggagcttgta tatccatttt cggatctgat
     2281 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     2341 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     2401 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     2461 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     2521 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2581 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     2641 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     2701 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     2761 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     2821 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     2881 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     2941 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3001 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3061 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3121 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3181 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3241 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3421 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5581 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     5641 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     5701 gttgga
//
