LOCUS       sequence-257008-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EcSSPB-nano (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-micro
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     719..738
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        719..737
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(793..999,1000..1002,1003..1122)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein and with the mutation R74Q to reduce the 
                     affinity for iLID"
                     /label=SspB micro
                     /note="dimerizes with iLID at moderate affinity to create 
                     iLID micro (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNAQFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     primer_bind     complement(1153..1170)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1159..1383
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1429..1857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1516..1535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1726..1747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1866..1886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1871..2200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2051..2186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2113..2132
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2267..3061
                     /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(2321..2340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2931..2950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3235..3356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3272..3291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3326..3345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3405..3421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3405..3421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3418..3440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3429..3445
                     /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(3453..3459,3460..3477,3478..3483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3498..3519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3636..3653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3807..4395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3887..3906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4566..5357,5358..5426))
                     /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     5189..5208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5427..5531)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5606..5625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
       61 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      721 atacgactca ctatagggag acccaagctg accatggcca ccggtgaatt ctcaggctct
      781 ggatcccccg ggagctcccc gaaacgccct aagctgctgc gtgaatatta cgattggctg
      841 gttgataaca gctttacccc atatctggtg gtggatgcca catacctggg cgtgaacgtg
      901 cccgtggagt atgtgaaaga cggtcagatc gtgctgaatc tgtctgcaag tgcgaccggc
      961 aacctgcaac tgacaaatga ttttatccag ttcaacgccc agtttaaggg cgtgtctcgt
     1021 gaactgtata tcccgatggg tgccgctctg gccatttacg ctcgcgagaa cggcgatggt
     1081 gtgatgttcg aaccagaaga aatctatgac gagctgaata ttggtcccgg gtaagtttaa
     1141 accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1201 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1261 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1321 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     1381 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     1441 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1501 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1561 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1621 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1681 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1741 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1801 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     1861 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1921 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1981 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2041 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2101 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2161 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctctcgg gagcttgtat
     2221 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2281 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2341 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2401 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2461 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2521 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2581 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2641 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2701 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2761 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     2821 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2881 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2941 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3001 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3061 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3121 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3181 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3241 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3301 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3361 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3421 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3481 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3541 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3601 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4561 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4621 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4681 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4741 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4801 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4861 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4921 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4981 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5041 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5101 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5161 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5221 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5281 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5341 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5401 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5461 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5521 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     5581 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     5641 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     5701 caacaa
//
