LOCUS       sequence-13014-e        7096 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 7096)
  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 7096)
  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-2-mEGFP
FEATURES             Location/Qualifiers
     source          1..7096
                     /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      2245..2254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2251..2253,2254..2256,2257..2970)
                     /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(2296..2317)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2557..2576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3016..3033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3022..3246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3530..3577
                     /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             3585..4607
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3585..4607
                     /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    4737..4858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4774..4793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4828..4847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /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(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /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     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7033)
                     /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 taccatggga
      961 gaggacgctg cacaggccga aaagttccag caccctgggt ctgacatgcg gcaggaaaag
     1021 ccctcgagcc ccagcccgat gccttcctcc acaccaagcc ccagcctgaa cctagggaac
     1081 acagaggagg ccatccggga caactcacag gtgaacgcag tcacggtgct cacgctcctg
     1141 gacaagctgg tgaacatgct agacgctgtg caggagaacc agcacaagat ggagcagcga
     1201 cagatcagtt tggagggctc cgtgaagggc atccagaatg acctcaccaa gctctccaag
     1261 taccaggcct ccaccagcaa cacggtgagc aagctgctgg agaagtcccg caaggtcagc
     1321 gcccacacgc gcgcggtcaa agagcgcatg gataggcagt gcgcacaggt gaagcggctg
     1381 gagaacaacc acgcccagct cctccgacgc aaccatttca aagtgctcat cttccaggag
     1441 gaaaatgaga tccctgccag cgtgtttgtg aaacagcccg tttccggtgc cgtggaaggg
     1501 aaggaggagc ttccggatga aaacaaatcc ctggaggaaa ccctgcacac cgtggacctc
     1561 tcctcagatg atgatttgcc ccacgatgag gaggccctgg aagacagtgc cgaggaaaag
     1621 gtggaagaaa gtagggcaga gaaaataaaa agatccagcc tgaagaaagt ggatagcctc
     1681 aagaaagcat tttctcgcca gaacatcgag aaaaagatga acaagctggg gacaaagatc
     1741 gtatctgtag agaggagaga gaagattaag aaatctctca cgtcaaatca ccagaaaata
     1801 tcctcaggaa aaagctcccc cttcaaggtt tctcccctca ctttcgggcg gaagaaagtc
     1861 cgagagggag aaagccatgc agaaaatgag accaagtcag aagacctgcc tagcagtgag
     1921 cagatgccaa atgaccagga agaggagtcc tttgcagagg gtcattccga agcgtccctc
     1981 gccagcgctc tggtggaagg ggaaattgca gaggaggctg ctgagaaggc gacctccagg
     2041 gggagtaact cggggatgga cagcaacatc gacttgacta ttgtggaaga tgaagaggag
     2101 gagtcagtgg ccctggaaca ggcacagaag gtacgctatg agggtagcta cgcgctaaca
     2161 tccgaggagg cggagcgctc cgatggggac cccgtgcagc ccgccgtgct ccaggtgcac
     2221 cagacctccg cggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     2281 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2341 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2401 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2461 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2521 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2581 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2641 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2701 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2761 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2821 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2881 gaccccaacg agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc
     2941 actctcggca tggacgagct gtacaagtaa agcggccgct cgagtctaga gggcccgttt
     3001 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3061 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3121 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3181 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3241 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat
     3541 tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca
     3601 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     3661 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     3721 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     3781 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     3841 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     3901 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     3961 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4021 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4081 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4141 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     4201 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     4261 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     4321 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     4381 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     4441 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     4501 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     4561 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag
     4621 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4681 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4861 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7081 tcccgatccc ctatgg
//
