LOCUS       sequence-256984-        5403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector containing A-B-C casette with linker L4 in site B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5403)
  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 5403)
  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_L4(EAAAK)3
FEATURES             Location/Qualifiers
     source          1..5403
                     /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"
     primer_bind     complement(891..908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    897..1121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1167..1595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1254..1273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1464..1485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1604..1624)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1609..1938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1789..1924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1851..1870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2005..2799
                     /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(2059..2078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2669..2688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2973..3094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3064..3083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3143..3159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3143..3159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3156..3178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3167..3183
                     /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(3191..3197,3198..3215,3216..3221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3236..3257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3374..3391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3545..4133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3625..3644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4304..5095,5096..5164))
                     /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     4927..4946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5165..5269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5344..5363)
                     /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 tcgaagcggc ggcgaaagaa gcggcggcga
      841 aagaagcggc ggcgaaagga tcccccgggt aagtttaaac cgctgatcag cctcgactgt
      901 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
      961 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1021 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1081 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1141 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1561 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1621 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1681 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1741 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1801 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1861 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1921 ggcctaggct tttgcaaaaa gctctcggga gcttgtatat ccattttcgg atctgatcaa
     1981 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2041 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2101 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2161 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2221 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2281 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2341 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2401 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2461 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2521 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2581 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2641 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     2701 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     2761 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     2821 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2881 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     2941 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3061 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3121 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3181 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3241 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3301 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4981 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5161 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5221 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5281 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5341 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5401 gga
//
