LOCUS       sequence-138521-        8188 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 8188)
  AUTHORS   Erdmann KS, Mao Y, McCrea HJ, Zoncu R, Lee S, Paradise S, Modregger 
            J, Biemesderfer D, Toomre D, De Camilli P
  TITLE     A role of the Lowe syndrome protein OCRL in early steps of the 
            endocytic pathway.
  JOURNAL   Dev Cell. 2007 Sep . 13(3):377-90.
  PUBMED    17765681
REFERENCE   2  (bases 1 to 8188)
  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: "Dev Cell. 
            2007 Sep . 13(3):377-90."
COMMENT     Sequence Label: mRFP-human APPL1
FEATURES             Location/Qualifiers
     source          1..8188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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)."
     primer_bind     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    218..339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(229..248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     283..302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(513..1307)
                     /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(624..643)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1234..1253
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1374..1703)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1388..1523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1442..1461)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1688..1708
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1717..2145)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1827..1848)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2039..2058
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2191..2415
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2404..2421
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2441..2459
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2441..2458
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4623..5294)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     primer_bind     complement(4682..4705)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4880..4899)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5149..5167
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5291..5300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5318..5337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5319..5337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5382..5585)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5411..5431)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5586..5965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6137..6156
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6231..6335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6336..6404,6405..7196)
                     /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     complement(6554..6573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7367..7955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7856..7875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8109..8126
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      181 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
      241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      301 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
      361 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
      421 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
      481 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
      541 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
      601 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
      661 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
      721 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
      781 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
      841 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
      901 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
      961 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     1021 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     1081 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     1141 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     1201 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     1261 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     1321 catcctgtct cttgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa
     1381 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
     1441 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     1501 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     1561 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     1621 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     1681 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
     1741 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1801 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1861 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1921 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1981 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2041 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2101 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag
     2161 agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc
     2221 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     2281 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     2341 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     2401 aactagaagg cacagtcgag gctgatcagc gagctctagc atttaggtga cactatagaa
     2461 tagggccctc tagatgcatg ctcgagctat gcttctgatt ctctcttctt tcctccttct
     2521 cccaaatcac tctcttctga ctggctactg ctaaggacaa caaactgccc ctcactactg
     2581 gcttggtttg gtctactgga agcagctatc aaccgacttt gttcttccaa gtccttttca
     2641 atctgttttt gtttattgag ttctttctgt tgcttctctt ttactctctc tatttctttt
     2701 tgtttttctg atgccctacg atccagttca gcatgcaaag ctatctgttt tgccagtcca
     2761 acagaatcac atatcttttc cccctcattg tttgactcaa atatatagca gactgatgac
     2821 agattacttt cacttctccc gcttgatgtc cgaagaacaa atccaaaaag gcgcttattt
     2881 tcctggtgtg tagcatacaa aactacacaa ggtaatggaa acgtgagcct tgtaacttgt
     2941 gtctgtggat caattaactt taaacagtca caagtgacta ataaatgcga ttctgtcata
     3001 cgaaagatgt tatggatggc ccgggcagct aagatttggc gcatagtttc ataaacaaca
     3061 tctggatggt catctgattt cacctccatt gaaccaagga atcggacaat aaataactga
     3121 tgaagaatag aatcttcagt ttcagatttt gtacttcctc cagattctcc aaatggattt
     3181 gtacgcctgc ctccctggcc aaaggctttt gcctggccag gctgatcttc acacacagga
     3241 gaaattatgt caaactgtat tggggtgtct ggggcaacaa gagaatctag agagagggca
     3301 gccaaatttg tagactcaga tcctaaggat cctgaactgc tggttcgagc tgtcggtggc
     3361 cgagattgtc ctgctgctgg ccgcaggctc tcgtgcctct gctggaaaga tggggaagga
     3421 gtgacagctt ccagagctga ttgatttact cgtgcagcag tttcctctgg attttcactt
     3481 aagtatattt gtttagatat gttgtttatt gtacagatcc actcttcatg atctttttta
     3541 ctctctgctt gcaaaattga agattttttt ccatcgaaag aggtgatctg aaaacaatat
     3601 cgtctgtctt cacagtccac agccatcact gaacagttgt ctatgtccat ggccaggcct
     3661 cctgctacat ccccacgggc ctgactcatt aaatttccac cctgcgtgaa gtaaaactgt
     3721 ctgtcccagg tagatgacac caagcctgtt ttattcctag cattaaggta tccagccttt
     3781 cgggttaaat ttcgattaac aggaaatttg gtggggtctg ggtcaggcac atataaggga
     3841 tcactggcta cttccaaatc ctctattgtc tgttgcatgg tctctatatc actgtccatt
     3901 tccctgcgaa cattctgaac gcttgttcca atattagcta aaaattcttc cagttgttca
     3961 ttaagatttt cagaacccat cttaaagaaa cttatctgag cttgcatgta cccaagtaga
     4021 ggttctaaca atgctatttt cttcttgtac tgaagagtat ttaatgcaca aaaataatgc
     4081 atcatggtct ggtgttgttt ctttctggat gtgtacacat cttctgttac ttcatacttc
     4141 accttgtcat tttctctttt ttttgataaa cggctatatc tattaatcgc agcatcatga
     4201 tcattacttg caatctgaaa tacttccttt aatgttagta tttctttcag atctctttct
     4261 ttaaactggg taatggggaa catcatggca tcagcaagtt gagttgaaag cactgcatga
     4321 caagagctaa gctcatctat aacttttgaa aactgttgca atgtagagct cataacttca
     4381 tcatcacctc ccaatggaaa acgctgtttt tcatattctt ttaaaagttt tgaggtcagg
     4441 tgtgttgctg cacttaattc attctgtgca tcataaatcc gatgcatagc ttgatacaac
     4501 tggttcatat agttggaaat agctgtggca tcttcttcaa atacacctag taaagacctt
     4561 gtctgcgggc tgtcctccag cgtctcctcg atgggcagct tgtcgatccc cggcatggta
     4621 ccggcgccgg tggagtggcg gccctcggcg cgctcgtact gttccacgat ggtgtagtcc
     4681 tcgttgtggg aggtgatgtc cagcttgatg tcggtcttgt aggcgccggg cagctgcacg
     4741 ggcttcttgg ccatgtaggt ggtcttgacc tcggcgtcgt agtggccgcc gtccttcagc
     4801 ttcagcctca tcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggtg
     4861 gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg
     4921 ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg
     4981 gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag
     5041 gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac
     5101 tggaactgag gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc
     5161 agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac
     5221 tcgtggccgt tcacggagcc ctccatgcgc accttgaagc gcatgaactc cttgatgacg
     5281 tcctcggagg ccatggtggc gaccgaagct tgggtctccc tatagtgagt cgtattaatt
     5341 tcgataagcc agtaagcagt gggttctcta gttagccaga gagctctgct tatatagacc
     5401 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     5461 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     5521 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     5581 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     5641 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     5701 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     5761 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     5821 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     5881 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     5941 taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc
     6001 gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt
     6061 gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga
     6121 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca taggggatcg
     6181 ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7501 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8161 tccccgcgcg ttggccgatt cattaatg
//
