LOCUS       sequence-489670-        5935 bp    DNA     circular SYN 17-OCT-2025
DEFINITION  Express human CCDC32 in mammalian cells with HA tag at C terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  AUTHORS   Wan C, Puscher H, Ouyang Y, Wu J, Tian Y, Li S, Yin Q, Shen J
  TITLE     An AAGAB-to-CCDC32 handover mechanism controls the assembly of the 
            AP2 adaptor complex.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2409341121. doi: 
            10.1073/pnas.2409341121. Epub 2024 Aug 15.
  PUBMED    39145939
REFERENCE   2  (bases 1 to 5935)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-08-20"; volume: "121(34):e2409341121. 
            doi"; pages: " 10.1073/pnas.2409341121. Epub 2024 Aug 15"
COMMENT     Sequence Label: pcDNA3.1-hCCDC32-HA
FEATURES             Location/Qualifiers
     source          1..5935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1471..1497
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1526..1543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1532..1756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1802..2230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1889..1908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2099..2120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2239..2259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2244..2573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2424..2559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2486..2505
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2640..3434
                     /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(2694..2713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3304..3323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3608..3729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3645..3664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3699..3718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3778..3794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3778..3794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3791..3813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3802..3818
                     /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(3826..3832,3833..3850,3851..3856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3871..3892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4009..4026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4180..4768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4260..4279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4939..5730,5731..5799))
                     /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     5562..5581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5800..5904)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 ctagccacca tgaaaatgtt tgagagcgct gattcaactg caactagatc tggccaggat
      961 ctctgggctg aaatttgttc ctgtctgcca aatcctgaac aagaagatgg tgccaacaat
     1021 gcattctcag actcctttgt ggattcttgc cctgaaggtg aaggccagag ggaggtggct
     1081 gactttgctg tccagccagc tgtaaagcct tgggctccct tgcaggattc agaggtatac
     1141 ttggctagtc tagagaagaa gctaagaaga atcaaaggtt taaatcagga agtgacttcc
     1201 aaggacatgc ttcgaactct ggcccaagcc aagaaggaat gctgggatcg gttcctccag
     1261 gagaagttag cttcagagtt ctttgtggat ggacttgatt ctgatgagag caccttggaa
     1321 catttcaaga ggtggctcca gccagataaa gtagccgtca gcacagagga ggtccagtat
     1381 ctgattcctc cagagtcaca ggttgagaag ccagtggccg aggacgagcc agcagccggg
     1441 gacaagccag cagcagcaga acagctcgag tacccatacg atgttccaga ttacgcttga
     1501 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1561 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1621 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1681 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1741 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     1801 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1861 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1921 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1981 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2041 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2101 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2161 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2221 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2281 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2341 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2401 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2461 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     2521 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     2581 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     2641 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2701 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2761 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2821 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2881 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2941 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3001 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3061 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3121 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3181 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3241 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3301 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3361 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3421 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3481 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3541 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3601 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3661 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3721 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3961 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4021 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4081 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4141 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4201 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4261 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4321 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4381 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4441 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4501 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4561 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4621 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4681 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt tttttngttt gcaagcagca
     4741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
