LOCUS       sequence-13013-e        6998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6998)
  AUTHORS   Hayer A, Stoeber M, Bissig C, Helenius A
  TITLE     Biogenesis of caveolae: stepwise assembly of large caveolin and 
            cavin complexes.
  JOURNAL   Traffic. 2010 Mar . 11(3):361-82.
  PUBMED    20070607
REFERENCE   2  (bases 1 to 6998)
  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: "Traffic. 
            2010 Mar . 11(3):361-82."
COMMENT     Sequence Label: cavin-1-mEGFP
FEATURES             Location/Qualifiers
     source          1..6998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        203..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    788..806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    809..827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     837..861
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2147..2156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2153..2155,2156..2158,2159..2872)
                     /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(2198..2219)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2459..2478)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2918..2935)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2924..3148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3281..3300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3432..3479
                     /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             3487..4509
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3487..4509
                     /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
                     "
     polyA_signal    4639..4760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4676..4695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4730..4749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4809..4825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4809..4825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4822..4844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4833..4849
                     /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(4857..4863,4864..4881,4882..4887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4902..4923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5040..5057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5211..5799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5291..5310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5970..6761,6762..6830))
                     /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     6593..6612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6831..6935)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
       61 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      121 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      181 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      241 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
      841 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      901 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tcgccatgga
      961 ggatgtcacg ctccatatcg ttgagcggcc gtattccgga tttcccgatg cttcctcaga
     1021 gggcccggag cccacccaag gggaggcgcg ggccacggag gagccgtcgg ggaccggctc
     1081 cgacgagctg atcaagtcgg accaggtgaa cggtgtgctg gtgctgagcc ttctggataa
     1141 aatcatcggc gccgttgacc agatccagct gacccaagcc cagctggagg agcgacaggc
     1201 ggagatggag ggcgctgtgc agagcatcca gggagagctg agcaagctgg gcaaggcgca
     1261 cgccaccacg agcaacaccg tgagcaagtt gctggagaag gtgcgcaagg tcagcgtcaa
     1321 cgtgaagacc gtgcgcggca gcctggagcg ccaggccggc cagataaaga aactggaggt
     1381 caacgaggcg gagctgctga ggcgccgcaa cttcaaagtc atgatctacc aggatgaagt
     1441 caagctgccg gccaaactga gcgtcagcaa gtcgctgaaa gagtcggagg cactgcctga
     1501 gaaggagggt gacgagctgg gcgagggcga gcggcccgag gatgacaccg cggcgatcga
     1561 gctgtcgtcc gacgaggcgg tggaggtgga ggaggtgatc gaggagtccc gcgccgagcg
     1621 catcaagcgc agcggcctgc ggcgcgtgga cgacttcaag aaggccttct ccaaggagaa
     1681 gatggagaag accaaggtgc gcacgcgtga gaacctggag aagacgcgcc tgaagaccaa
     1741 ggagaacctg gagaagacac ggcacacgct ggagaagcgc atgaacaagc tgggcacgcg
     1801 cctggtgccc gtggagcgac gagagaagct gaagacatcc cgggacaagc tgcgcaagtc
     1861 cttcacgccc gaccatgtgg tgtatgcgcg ctccaagacc gctgtctaca aggtgccgcc
     1921 tttcaccttc cacgtcaaga agatccgcga gggcgaggtg gaggtgctga aggccaccga
     1981 gatggtggag gtgggtcccg aggacgacga ggttggcgcg gagcgcggcg aggccactga
     2041 cctgctgcgc gggagcagcc ccgacgtgca cacgctgctg gagatcaccg aggagtcgga
     2101 cgccgtcctg gtggacaaga gcgacagcga cgcggatcca ccggtcgcca ccatggtgag
     2161 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2221 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2281 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2341 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2401 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2461 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2521 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2581 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2641 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2701 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2761 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tccttaagga
     2821 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     2881 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2941 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3001 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3061 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3121 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3181 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3241 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3301 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3361 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3421 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     3481 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     3541 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     3601 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     3661 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     3721 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     3781 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     3841 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     3901 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     3961 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4021 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4081 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4141 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4201 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4261 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4321 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4381 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4441 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4501 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4561 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4621 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4681 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4741 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4801 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4861 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4921 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5041 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5101 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5401 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5641 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5881 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5941 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6001 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6061 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6121 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6181 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6241 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6301 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6361 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6421 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6481 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6541 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6601 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6661 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6721 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6781 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6841 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6961 gacgtcgacg gatcgggaga tctcccgatc ccctatgg
//
