LOCUS       sequence-342782-        5019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses protein with one aa substitution to affect potential 
            phosphorylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5019)
  AUTHORS   Marchak A, Grant PA, Neilson KM, Datta Majumdar H, Yaklichkin S, 
            Johnson D, Moody SA
  TITLE     Wbp2nl has a developmental role in establishing neural and 
            non-neural ectodermal fates.
  JOURNAL   Dev Biol. 2017 Sep 1;429(1):213-224. doi: 
            10.1016/j.ydbio.2017.06.025. Epub 2017 Jun 27.
  PUBMED    28663133
REFERENCE   2  (bases 1 to 5019)
  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"; doi: 
            "10.1016/j.ydbio.2017.06.025"; journalName: "Dev Biol"; date: 
            "2017-09-1- 1"; volume: "429"; issue: "1"; pages: "213-224"
COMMENT     Sequence Label: pCS2+-wbp2nlT45A
FEATURES             Location/Qualifiers
     source          1..5019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1071..1205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1095..1114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1149..1168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1318..1338)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1318..1336)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1357..1373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1357..1373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1370..1392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1381..1397
                     /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(1405..1411,1412..1429,1430..1435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1450..1471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1588..1605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1759..2347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1839..1858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2518..3309,3310..3378))
                     /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     3141..3160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3379..3483)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3509..3964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3646..3667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3858..3877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4035..5019
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttaaagcaac tccattccag caattactga
      121 gtagtgcgca gggagaagtt gaaggcttta gtgcgagatg tcactcaata aggaccactc
      181 tcagggggcc gggatcattg tcagcaacgg ggagagtatt ctcagacagt gtaaagatgt
      241 ggaactgtcc ttcagcgata tgtctcacaa atcagaagca ttcagaggcg caaagaaagg
      301 atcgctattt ctcaccgctt acagggtgat cttcctaaac aaggggaagg atcccatgat
      361 gtcctttaat atgccattct acatgatgaa aggatgctcc atagaacagc cagtgttctc
      421 tgccaactac atcaagggaa ccatcagtgc tgaacctgga ggtggctggg agggtcaagc
      481 ctcttttaaa attacattta atagtggagg tgctattgaa tttggacaaa tcatgttcaa
      541 aatggcaact tgtgcttcca gaacccctcc tgttcctcat gctgcatatg gatacacacc
      601 tgctccagga ggctatgcac atggcgggta ccctccagct ccaggaatat atacaccccc
      661 accccaacca tcaggccctt atccatatgg accccctgct atgaatggat atggaccacc
      721 tccaaatcct atgggatacc cgtatgctcc accagcagga cctggcatgt atccaccccc
      781 tcctgaaatg aatcccattt acatggctcc tccaccacct tatcctggac ccccctacaa
      841 tgggaccccc tacaatggaa ccccagcacc ttctgctccc actggatgca tgaatgctgg
      901 tatgcctgga ggtagcaaag ctgcagaagc tgcatccagt gcgtattata acccagctga
      961 ccctcacaat gtctatatgc ctatggatcg tcctcctcca tacgctccaa atgatgataa
     1021 aaagaacaat taactcgagc ctctagaact atagtgagtc gtattacgta gatccagaca
     1081 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1141 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1201 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1261 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     1321 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1381 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     1441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1621 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2581 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2641 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2701 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2761 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2821 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2881 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2941 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3001 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3061 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3301 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3361 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3421 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3481 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     3541 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3601 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3661 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3721 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3781 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3841 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3901 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3961 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4021 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
     4081 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     4141 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     4201 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     4261 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     4321 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     4381 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     4441 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     4501 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     4561 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     4621 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     4681 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     4741 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     4801 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
     4861 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     4921 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     4981 taatgggagg tctatataag caatgctcgt ttagggaac
//
