LOCUS       sequence-139173-        6059 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 6059)
  AUTHORS   Allaire PD, Seyed Sadr M, Chaineau M, Seyed Sadr E, Konefal S, 
            Fotouhi M, Maret D, Ritter B, Del Maestro RF, McPherson PS
  TITLE     Interplay between Rab35 and Arf6 controls cargo recycling to 
            coordinate cell adhesion and migration.
  JOURNAL   J Cell Sci. 2013 Feb 1;126(Pt 3):722-31. doi: 10.1242/jcs.112375. 
            Epub 2012 Dec 21.
  PUBMED    23264734
REFERENCE   2  (bases 1 to 6059)
  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: "J Cell 
            Sci."; date: "2013-02-1"; volume: "126(Pt 3):722-31. doi"; pages: " 
            10.1242/jcs.112375. Epub 2012 Dec 21"
COMMENT     Sequence Label: Myc-Rab35 S22N
FEATURES             Location/Qualifiers
     source          1..6059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(565..1359)
                     /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(676..695)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1286..1305
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1426..1755)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1440..1575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1494..1513)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1740..1760
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1769..2197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2091..2110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2456..2473
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2493..2511
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2493..2510
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3219..3228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3241..3260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3242..3260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3305..3508)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3334..3354)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3509..3888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4060..4079
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4154..4258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4259..4327,4328..5119)
                     /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(4477..4496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5290..5878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5779..5798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6032..6049
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
      541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
      601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
      661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
      721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
      781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
      841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
      901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
      961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
     2521 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cttagcagca
     2581 gcgtttcttt cgtttactgt tcttcgtgag cttcaccaca tcgttctgtt gttgctgctg
     2641 ctgttttgcc aggttgtctt tctttgctcg gaggaccagc tccgtgatgc agttgaacat
     2701 ctcttccacg ttgacattct ccttggcgct ggtctcgaac aactggatgc ccatctgccc
     2761 ggcgaatttg taggcatctt ccgtctccac caccttccgc tcagggtcgt cattcttatt
     2821 acccactaat attcggcaca catcatcaca gttctggttg atttcgtgaa gccaccgctt
     2881 gacgttgaca aaggactcgg cactggtgac gtcgtaaacc acaatgaccc cgtgggtccc
     2941 ccgataatac gtggaggtga tggtgcggaa gcgctcctgc cccgctgtgt cccagatctg
     3001 cagcttcacc ttctccccgt tgatctccac ggtccggatc ttgaaatcca ctccgatcgt
     3061 ggtgatgtag ctgcctgaga aagtgttgtc tgcaaaacgc aacagtaaac tgttcttgcc
     3121 cacaccgctg tcgccgatga tgagcagctt gaagaggtgg tcgtagtccc gggccatgga
     3181 tcccaggtcc tcctctgaga tcagcttctg cattgatgcc atggtggcaa gcttgggtct
     3241 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
     3301 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     3361 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     3421 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     3481 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     3541 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     3601 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     3661 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     3721 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     3781 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     3841 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
     3901 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     3961 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     4021 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     4081 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     4141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4201 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4261 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4321 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4381 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4441 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4501 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4561 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4621 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4681 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4741 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4801 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4861 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4921 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4981 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5041 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5101 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5401 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5941 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6001 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
