LOCUS       sequence-102902-        6277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KCTD13 N-terminus disruption by homolgy-directed repair.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  AUTHORS   Martinez RA, Stein JL, Krostag AR, Nelson AM, Marken JS, Menon V, 
            May RC, Yao Z, Kaykas A, Geschwind DH, Grimley JS
  TITLE     Genome engineering of isogenic human ES cells to model autism 
            disorders.
  JOURNAL   Nucleic Acids Res. 2015 Mar 11. pii: gkv164.
  PUBMED    25765640
REFERENCE   2  (bases 1 to 6277)
  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: "Nucleic 
            Acids Res. 2015 Mar 11. pii: gkv164."
COMMENT     Sequence Label: AI-CN590
FEATURES             Location/Qualifiers
     source          1..6277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    526..559
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        600..811
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             824..1092
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             join(1126..1128,1129..1131,1132..1842)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1171..1192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1432..1451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1779..1800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1852..1908
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2601..2620)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2666..2721
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2720..2739)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    3056..3089
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3660..3676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3660..3676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3673..3695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3684..3700
                     /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(3708..3714,3715..3732,3733..3738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3753..3774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3891..3908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4062..4650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4142..4161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4821..5612,5613..5681))
                     /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     5444..5463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5682..5786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5854..5872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5910..5932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6032..6051
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6245..6267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6259..6276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6260..6276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgagct cggtacctcg cgaatgcatc tagatttggg gccggccttc cgtaagtgtt
       61 ggttattgct gggggtaaag gattggggga ggagaagaga aattatgaat gagaaagggg
      121 aggagatggg atgtgggggc tggagggagc tgagaatgag ttgactggac agtagcgggc
      181 agagagaagg ggcatgagga gccccggtgg tgagagactg cggggaacgg gagttggggg
      241 tctggggtgg gggcacggta ggggcgccgc tcgtacctcc ggcatcggtc agcacctcca
      301 cgcggccgct gaacatggct ttgagcatgg tgtcctgtcc cgtgagggtg cgcagcgtgg
      361 tgtagtgcaa cgagccgccc acgttcagct tcacgtattt gctgttcggg gtcagcggct
      421 tgagaccgta ggcggcgggg ccaggctcga gacccgaggg cttgggggct tccagggacg
      481 gggccgcggc ggcagccggg cccgaggcct ccgccgaggc gcgccataac ttcgtataat
      541 gtatgctata cgaagttatg cccgggcaag gatctgcgat cgctccggtg cccgtcagtg
      601 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      661 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      721 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      781 ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc
      841 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
      901 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
      961 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     1021 cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg
     1081 ccgttacaga tccaagctgt gaccggcgcc tacctcgagc ccaccatggt gagcaagggc
     1141 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1201 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1261 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1321 acctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1381 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1441 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1501 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1561 gccatcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1621 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1681 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1741 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1801 accgccgccg ggatcactct cggcatggac gagctgtaca agggaagcgg agctactaac
     1861 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctac tagtatgact
     1921 gaatacaaac ccactgtccg actggcaaca agagatgatg tcccaagagc cgtgagaaca
     1981 ctggcagcag ctttcgccga ctacccagcc acaagacaca ctgtggaccc cgatcggcat
     2041 atcgagagag tgaccgagct gcaggaactg ttcctgacaa gggtgggcct ggacattggg
     2101 aaggtctggg tcgcagacga tggagcagct gtggctgtct ggaccacacc agagagcgtg
     2161 gaagccggag ctgtctttgc agagatcggc cctcgaatgg cagaactgag cggctcccgg
     2221 ctggcagcac agcagcagat ggaggggctg ctggccccac accgccctaa ggaaccagca
     2281 tggttcctgg ccactgtggg agtctctccc gatcatcagg gcaaagggct gggaagtgct
     2341 gtggtcctgc ctggggtgga ggctgcagaa cgagctggag tcccagcatt tctggagaca
     2401 tctgccccca ggaacctgcc tttctatgaa cgcctgggct ttactgtgac cgccgatgtg
     2461 gaagtgcccg agggaccccg aacctggtgt atgacccgaa agcctggagc ctaatagacc
     2521 ggtactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     2581 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     2641 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     2701 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     2761 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     2821 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     2881 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     2941 ttttctttaa catccctaaa attttcctta cctgttttac tagccagatt tttcctcctc
     3001 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatcatt taaatataac
     3061 ttcgtataat gtatgctata cgaagttatg ctagcctatt acctactact tattattaat
     3121 taacatgccg ggtagcagcg gcggacggcg atcccaggat ctctccgcgc cctgcggcct
     3181 gctcccgaag accctcggcc ggcccccagc ccttgggcca gaccgctcgg cgcacacgcc
     3241 cactcaccgc agctactctg caagaccggc cctccgcccc atctcgctcg caccacccgg
     3301 aagccggcgc cgggcagctg cgcaggcgcg gccccgcccc tgaatggggg cggggctgcg
     3361 agtatggagc atgcgcagtg gcctcgctgg gggatcccgt agaggtttgg cgtgtggcct
     3421 gcgggttccc cgggcgggtc gacgtgcgct cctctctggt ctcgagtcac caggaccaga
     3481 tcgaggatgc ggccgacccc ctgcccgatg tggaaagcga agtctccacc tcgtgattgg
     3541 gtctctgctg tcagagagct tcacgagctt gaggggaaac agacagagag gtcgcggccg
     3601 caatcggatc ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg
     3661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5821 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5881 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5941 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6001 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6061 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6121 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6181 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6241 ttttcccagt cacgacgttg taaaacgacg gccagtg
//
