LOCUS       40924_4414        6544 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAMCRE1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  AUTHORS   Lu Y.
  TITLE     Establishing a novel L-ornithine self-sufficient cell factory with 
            enhanced production of fusarinine C through metabolic engineering in
            Aureobasidium melanogenum HN6.2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6544)
  AUTHORS   Lu Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2018) Ocean University of China, College of Marine
            Life, 5 Yushan Road, Qingdao, Shangdong 266003, China
REFERENCE   3  (bases 1 to 6544)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6544)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2018) Ocean University of China, College of Marine Life, 5 
            Yushan Road, Qingdao, Shangdong 266003, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2810..3838
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     CDS             3842..3862
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     terminator      complement(3871..4118)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     polyA_signal    complement(4136..4310)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4354..5376)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     primer_bind     complement(6150..6166)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcttga caatgagaaa agaccacctc ggaacgagga
     2281 tatacagcgg tatagatggc caaggacaga cttcaactgc ggctgtaggt agagggtcaa
     2341 agcagcattt caggacctaa ggagcttcaa gtagtgacgg tgcggagaaa cacggggaga
     2401 aacgggaagg aacgagaacc aggaagaaga aggaggggtt gtttgcgggg cggttcggga
     2461 catgggccaa gttgaagaga catcatggta tagacgcgta gaatgacgag actcccgaat
     2521 cgaagcgaaa gagaggctat ctgggccatc aaatggctcg ttatttgcaa gtgatggaag
     2581 ctgcaaaacg ccaagacatg tttcggcgag cggggctggc ttgagcggcg agctttcgtt
     2641 cattattcac ccaacagttt tttaagtcat cctttataat cctcactcct ccccctcgtc
     2701 cgactttgta accactccat aacacccatt cacagtaggt cccccatcac catctcaact
     2761 ccccctcctc catcttcaaa gctctaacaa ctaccccgca gacacaaaca tgtctaacct
     2821 cctcaccgtc caccaaaacc ttccagctct tccagttgac gctacttctg acgaagtccg
     2881 taagaacctc atggacatgt tcagagacag acaggctttc tccgagcata cttggaagat
     2941 gctcctctcc gtttgcagat cttgggcagc ttggtgtaag ctcaacaaca gaaagtggtt
     3001 cccagcagaa ccagaagacg tcagagatta cctcctctac ctccaagcta gaggtttggc
     3061 agtcaagact atccagcagc atctcggtca actcaacatg ctccacagaa gatctggcct
     3121 tcctagacct tccgattcta acgccgtctc tcttgtcatg cgtagaatcc gtaaggagaa
     3181 cgttgacgca ggagaaagag ctaagcaagc tctcgctttc gagagaacag acttcgatca
     3241 ggtccgttct ctcatggaaa actccgatcg ttgccaggac atcagaaacc ttgctttcct
     3301 cggcatcgct tacaacactc ttctcagaat cgccgagatc gctagaatca gagtcaagga
     3361 catctctaga accgacggag gtagaatgct catccatatc ggccgtacta agactctcgt
     3421 ctctacagca ggagtcgaaa aggctctctc tcttggagtc actaagctcg tcgaacgttg
     3481 gatctcagtc tcaggagttg cagacgaccc taacaactac ctcttttgca gagtcagaaa
     3541 gaacggagtt gcagctcctt ccgctacttc tcagctctct actagagctc tcgagggtat
     3601 cttcgaagct actcatcgtc tcatctacgg agctaaggac gattcaggtc agagatacct
     3661 cgcttggtca ggtcattctg ctagagtcgg agcagctaga gatatggcta gagcaggagt
     3721 ctctatccca gagatcatgc aagcaggagg ttggactaac gtcaacatcg tcatgaacta
     3781 catccgcacc ctcgattcag agactggcgc tatggtcaga cttcttgaag acggcgatcc
     3841 tcctaagaag aagagaaagg tctaacggcc gcaaattaaa gccttcgagc gtcccaaaac
     3901 cttctcaagc aaggttttca gtataatgtt acatgcgtac acgcgtctgt acagaaaaaa
     3961 aagaaaaatt tgaaatataa ataacgttct taatactaac ataactataa aaaaataaat
     4021 agggacctag acttcaggtt gtctaactcc ttccttttcg gttagagcgg atgtgggggg
     4081 agggcgtgaa tgtaagcgtg acataactaa ttacatgacg tcgactaatt cgggggatct
     4141 ggattttagt actggatttt ggttttagga attagaaatt ttattgatag aagtatttta
     4201 caaatacaaa tacatactaa gggtttctta tatgctcaac acatgagcga aaccctatag
     4261 gaaccctaat tcccttatct gggaactact cacacattat tatggagaaa ctcgagcttg
     4321 tcgatcgaca gatcccggtc ggcatctact ctatttcttt gccctcggac gagtgctggg
     4381 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct
     4441 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     4501 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     4561 gtcaagacca atgcggagca tatacgcccg gagtcgtggc gatcctgcaa gctccggatg
     4621 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     4681 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     4741 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     4801 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     4861 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     4921 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     4981 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg cgaccggttg
     5041 tagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgaacggcg
     5101 ggagatgcaa taggtcaggc tctcgctaaa ctccccaatg tcaagcactt ccggaatcgg
     5161 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     5221 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
     5281 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
     5341 ctcgacagac gtcgcggtga gttcaggctt tttcatgttt gacggtgatg tatggaagat
     5401 tgagtgaagt acggttgaac tttctgaaga gaggttagaa agagaggaag gaagaaaaga
     5461 atgagcagat ccaagaaata gcaaagtgga gcccgctcaa gaggtcaaag cttaccagat
     5521 acgattgctt gacgaaccgt ggtatgaaga cggtgatgag aagagggtgc gggaggggtg
     5581 gcctttttga gtgtgtgtga gagggcgcac tgactaagcg agccaaaggg tctagcttcg
     5641 gttgccccgc caagctagtt tttctctctc actttttttg ctgcttcgtt gggagcccac
     5701 cgagcggagc cagcagaaaa aaaaatgatg taacggatga tgagccaact gtcgggatcc
     5761 attgtcattc gtgattctag tactgtgggt ggctgcgaaa ctccttcact caatgaatta
     5821 caatcatgat tcgtgattcg caccgatccc accagcctct cgagaaaagg ggacttgaac
     5881 attcatgatt cacgaccctg tgattcatga atgccatccg atccagaacc gacttgctac
     5941 tcacgactca gactgaattt ttaaatgccc tcatgatttg attcatgatc agcaatgccc
     6001 tcatgatttg attcatgatt cgctgatcac agcggtgaag aacgtaaccc attcacgatt
     6061 cagaaggcaa gtactgcttc ttgctcggct gctttgttgc tgccgaggcg ggctagcttg
     6121 cacaaagatc gaggatcggt aatgaattca ctggccgtcg ttttacaacg tcgtgactgg
     6181 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6241 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6301 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6361 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     6421 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6481 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6541 gcga
//
