LOCUS       sequence-217304-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Capsaspora owczarzaki"" expression vector (backbone) modified from
            the pCR2.1. It bears the promoter and terminator regions from the 
            endogenous Elongation Factor 1 alpha (EF1a) gene (CAOG_07807)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Parra-Acero H, Ros-Rocher N, Perez-Posada A, Kozyczkowska A, 
            Sanchez-Pons N, Nakata A, Suga H, Najle SR, Ruiz-Trillo I
  TITLE     Transfection of Capsaspora owczarzaki, a close unicellular relative 
            of animals.
  JOURNAL   Development. 2018 May 11. pii: dev.162107. doi: 10.1242/dev.162107.
  PUBMED    29752387
REFERENCE   2  (bases 1 to 5127)
  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: 
            "Development. 2018 May 11. pii: dev.162107. doi: 
            10.1242/dev.162107."
COMMENT     Sequence Label: pONSY
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1350..1369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1351..1369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1376..1393)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1376..1392)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1385..1407)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1533..1961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1620..1639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1830..1851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2305..3099
                     /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(2359..2378)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2969..2988
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3117..3185,3186..3977)
                     /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     complement(3335..3354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4148..4736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4637..4656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4890..4907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5024..5045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5060..5065,5066..5083,5084..5090)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5098..5114
                     /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)."
     primer_bind     5103..5125
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5122..5127,1..11)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5122..5127,1..11)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acagctatga ccatgattac gccaagcttg gtaccaaatg cacagttagc aacgaccgac
       61 aatcagcaac ctccaaactc tcagtttttt tcccccaatc gttgtcgatt tagcttttga
      121 tgctgcattt tgaaatagaa cgacaaccga atttgggaga gagcactctt ttacaggttt
      181 gtgaaaagaa aaaaaatcga acgtgacttg cttccatctt ccttctcgaa gccgatctcg
      241 gcccatcccc gcttgtcccc tcctcccctc ctcgcaatcc gctagacagt gtctctgcac
      301 actggattgc ggtgcgcggg ccatccgatg cccctttcct cggcctggag atggacgtga
      361 aagtggaaaa cgaaaaacgc gaaacagaaa catgtggttc tttttttttt ctttttcttt
      421 ttcgccttcg ggcttttctc tctcgcttcc tcttctccgc tctcttcttc gcgctctttt
      481 gcctcttgcg tgagatgctc gtcttgcaat cgtctgccac tgctgcgctg tcacgatcga
      541 tcgcatcgga caagccgttt ctgttttgaa atttagaacg ttctggaaaa caacaaaaaa
      601 ttgccttttt ttcacttttt tcttttttcg tttttttccc tcttcaacac ttcctccaac
      661 tgtcgttctg cgtttgttct tcgctctcgt cgtgcacact cggattagga tccattcatc
      721 cccatcccct tgcgcattgc caaaaacgag cgtcgaaacg cgaacgtcag cgtggggtgt
      781 tgacgccgaa ccccacgagt gttttttttt aaaatgctgt ttcatcgttt tcttccttct
      841 ccccacgtgc aatcctgatt taaataattg tcgctttttg atttggaaat gctcactctt
      901 ccattttttt attatttaac acacacagaa caaccttcac aggatcccgg gactagtgat
      961 atctgaattt tgtgtttgcc aagacacatt tgtttatttc aatcatttct aagaccaaaa
     1021 agggatttca tgatttttgt tgcttgcatt gaagtttttt ttggttgtca ttttttttgc
     1081 gttggttatg agcgcaatat attccattat ggcaagaaac acgacgagtt gtttttacag
     1141 tggagggatg agcaaggaat ggggaaatgg agaacaagtg gaatgaaatg gtttgacttt
     1201 gtgcggcgaa agtgttgcca ttttcaatca tgctgcgcct ggcacctaga tgcgtgtgat
     1261 tgctcattgc tagtgctgtt ctcaccgcgg aagctttcaa tccatcacac tggcggccgc
     1321 tcgagcatgc atctagaggg cccaattcgc cctatagtga gtcgtattac aattcactgg
     1381 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1441 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     1501 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     1561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1861 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc
     1981 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
     2041 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     2101 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     2161 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     2221 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa
     2281 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2341 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2401 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2461 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2521 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2581 ctattgggcg aagtgccggg gcaggatctc ctgtcatccc accttgctcc tgccgagaaa
     2641 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2701 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2761 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2821 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2881 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2941 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3001 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3061 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga
     3121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4861 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4921 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4981 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5041 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5101 tgagcggata acaatttcac acaggaa
//
