LOCUS       sequence-439382-        7762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Cd2ap-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7762)
  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"
COMMENT     Sequence Label: Cd2ap-GFP
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    735..856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(772..791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     826..845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(905..921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(905..921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(918..940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    929..945
                     /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(953..959,960..977,978..983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    998..1019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1136..1153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1307..1895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1387..1406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2066..2857,2858..2926))
                     /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     2689..2708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2927..3031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3106..3125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3297..3676
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3677..3880
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3831..3851
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3882..3900
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3903..3921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3931..3955
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5980..6000
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(6013..6015,6016..6018,6019..6732)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6058..6079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6319..6338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6776..6793)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6782..7006
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7139..7158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    7290..7337
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             join(7345..7762,1..605)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(7345..7762,1..605)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
       61 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
      121 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
      181 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
      241 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
      301 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
      361 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
      421 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
      481 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
      541 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
      601 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
      661 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
      721 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      841 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
      901 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
      961 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1021 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1081 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1141 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1201 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3061 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3121 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3181 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3241 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     3301 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3361 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3421 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3481 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3541 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3601 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3661 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3721 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3781 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3841 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca gtgatagaga
     3901 tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg
     3961 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     4021 gactctagcg tttaaactta agcttaggcc tgcatggttg actatattgt ggagtatgac
     4081 tatgatgctg tacatgatga tgaattaact attcgagttg gagaaatcat caggaatgtg
     4141 aaaaagctac aggaggaagg gtggctggaa ggagaactaa atgggagaag aggaatgttc
     4201 cctgacaatt tcgttaagga aattaaaaga gagacggaat tcaaggatga cagtttgccc
     4261 atcaaacggg aaaggcatgg gaatgtagca agtcttgtac aacgaataag cacctatgga
     4321 cttccagctg gaggaattca gccacatcca caaaccaaaa acattaagaa gaagaccaag
     4381 aagcgtcagt gtaaagttct ttttgagtac attccacaaa atgaggatga actggagctg
     4441 aaagtgggag atattattga tattaatgaa gaggtagaag aaggctggtg gagtggaacc
     4501 ctgaataaca agttgggact gtttccctca aattttgtga aagaattaga ggtaacagat
     4561 gatggtgaaa ctcatgaagc ccaggacgat tcagaaactg ttttggctgg gcctacttca
     4621 cctatacctt ctctgggaaa tgtgagtgaa actgcatctg gatcagttac acagccaaag
     4681 aaaattcgag gaattggatt tggagacatt tttaaagaag gctctgtgaa acttcggaca
     4741 agaacatcca gtagtgaaac agaagagaaa aaaccagaaa agcccttaat cctacagtca
     4801 ctgggaccca aaactcagag tgtggagata acaaaaacag ataccgaagg taaaattaaa
     4861 gctaaagaat attgtagaac attatttgcc tatgaaggta ctaatgaaga tgaacttact
     4921 tttaaagagg gggagataat ccatttgata agtaaggaga ctggagaagc tggctggtgg
     4981 aggggcgaac ttaatggtaa agaaggagta tttccagaca attttgctgt ccagataaat
     5041 gaacttgata aagactttcc aaaaccaaag aaaccaccac ctcctgctaa ggctccagct
     5101 ccaaagcctg aactgatagc tgcagagaag aaatattttt ctttaaagcc tgaagaaaag
     5161 gatgaaaaat caacactgga acagaaacct tctaaaccag cagctccaca agtcccaccc
     5221 aagaaaccta ctccacctac caaagccagt aatttactga gatcttctgg aacagtgtac
     5281 ccaaagcgac ctgaaaaacc agttcctcca ccacctccta tagccaagat taatggggaa
     5341 gtttctagca tttcatcaaa atttgaaact gagccagtat caaaactaaa gctagattct
     5401 gaacagctgc cccttagacc aaaatcagta gactttgatt cacttacagt aaggacctcc
     5461 aaagaaacag atgttgtaaa ttttgatgac atagcttcct cagaaaactt gcttcatctc
     5521 actgcaaata gaccaaagat gcctggaaga aggttgccgg gccgtttcaa tggtggacat
     5581 tctccaactc acagccccga aaaaatcttg aagttaccaa aagaagaaga cagtgccaac
     5641 ctgaagccat ctgaattaaa aaaagataca tgctactctc caaagccatc tgtgtacctt
     5701 tcaacacctt ccagtgcttc taaagcaaat acaactgctt tcctgactcc attagaaatc
     5761 aaagctaaag tggaaacaga tgatgtgaaa aaaaattccc tggatgaact tagagcccag
     5821 attattgaat tgttgtgcat tgtagaagca ctgaaaaagg atcacgggaa agaactggaa
     5881 aaactgcgaa aagatttgga agaagagaag acaatgagaa gtaatctaga gatggaaata
     5941 gagaagctga aaaaagctgt cctgtcttct cgcggccgcg aaaatttata ttttcaaggt
     6001 gaaggtacca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     6061 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     6121 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     6181 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     6241 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     6301 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     6361 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     6421 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     6481 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     6541 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     6601 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     6661 gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg catggacgag
     6721 ctgtacaagt aatagtgact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     6781 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6841 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6901 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6961 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     7021 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     7081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     7141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     7201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     7261 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     7321 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     7381 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     7441 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     7501 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     7561 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     7621 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     7681 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     7741 ggcccattcg gaccgcaagg aa
//
