LOCUS       Exported                7551 bp ds-DNA     circular SYN 01-MAY-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCas61
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7551)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, May 1, 2016 from SnapGene 3.1.2
            http://www.snapgene.com
COMMENT     >pAQ1_YFP
FEATURES             Location/Qualifiers
     source          1..7551
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        510..614
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             615..1475
                     /codon_start=1
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    1630..2249
                     /note="pBR322 ori"
     rep_origin      1646..2234
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2496..3080
                     /note="glpK_up"
     misc_feature    3081..3095
                     /note="glpk up extra"
     misc_feature    3146..3169
                     /note="LacOperator"
     protein_bind    3148..3167
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator (symmetric)"
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     gene            3176..3243
                     /note="lacUV5"
     -35_signal      3213..3218
                     /note="-35"
     -10_signal      3236..3242
                     /note="-10"
     misc_feature    3243..3277
                     /note="Lac Operator"
     misc_feature    3249
                     /note="Predicted Transcription Start Site"
     protein_bind    3252..3268
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     misc_feature    3297..3302
                     /note="RBS"
     CDS             3311..3376
                     /codon_start=1
                     /note="Translation start"
                     /translation="MGHHHHHHHHHHSSGHIEGRHM"
     CDS             3317..3346
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /note="10xHis"
                     /translation="HHHHHHHHHH"
     CDS             3359..3370
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     gene            3374..4125
                     /note="sll1330"
     misc_feature    4168..4225
                     /note="T7 terminator"
     misc_feature    4324..4358
                     /note="pMB2 Promoter"
     CDS             4388..5470
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDFSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    5045
                     /note="W220"
     promoter        5740..5768
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /note="Pc promoter"
                     /note="class 1 integron promoter"
     misc_feature    5740..5745
                     /note="-35 box"
     misc_feature    5760..5768
                     /note="-10 box"
     CDS             5957..6490
                     /codon_start=1
                     /note="gentamicin-3-acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    6757..7338
                     /note="glpk_dn"
     promoter        complement(7346..7364)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7371..7387)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7528..432
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
       61 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      181 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      301 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      421 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      481 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2341 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2401 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2461 attaatgcag ctggcacgac aggtttcccg actggagcga ttggctatga tctaccaaag
     2521 ctggcgattt ttctgcaaac cgaattagaa ccacaatttg attttttaaa gccgattacc
     2581 attgccaacc tgaaggaaat agcgatcgcc tttatggagg atggccacga ggcttgtcgg
     2641 gtctcccatt attgtggatc ggtaccggat gagcgggcga gctttaattt acgcttgcgg
     2701 caatatcgcc aggcccagcc ttggctgcga aatcatctcg atcctgcaag gggcgatcgc
     2761 ctgcaccatt ggtcggatca ccaacgcacc attttctacg gcaggcgcac caatcccgac
     2821 acccagcagc ggcttgtgtt agtggcgcac atggccgggg ctccgaagac cgttgagatt
     2881 ggcaaatggc tcgccctgga tttggatcgt tggcagttgg cgatcgccac accgactttg
     2941 aagatcaaca ccatctatga cttagcccaa attcacttgc acaatggcga aggttttctg
     3001 ttatctgaaa ttcctcccta aatgatgtct tttgagccta aaaacacact tttttgacct
     3061 aatttaaccc atttaaaaaa ctttatttaa taatgaccat ggcccatcaa aaatacattc
     3121 ttgcgttaga cctcggtacc acaaacgaat tgtgagcgct cacaattcgg aattcttaac
     3181 aaaaaagcag gaataaaatt aacaagatgt aacagacata agtcccatca ccgttgtata
     3241 aatgtgtgga attgtgagcg gataacaatt tcacacacca actcataaag tcaagtagga
     3301 gattaattcc atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat
     3361 cgaaggtcgt catatgaatc cagtctacat atcagtcgta gagggtaacc cccatctccg
     3421 ctctctattg agctggcacc tccagcagag cggctacctt gtccaacaat gtagcggttt
     3481 ccaccaagcc cgccaggcat ttaataacca attgcccacc ctggccgtga ttgattctga
     3541 ccttactgat ggggatggca ttgagctttg ccgttggtta tatcaacagc accagtcaat
     3601 gatttttatt ctttccgcca aggacacaga aaaagacatt gtccatgggt taaaggctgg
     3661 ggctgatgat tatctaacca agccctttgg catgcaggaa tttttagccc gcatagaatg
     3721 tctaattcgc cgggtccgca ctgtggccgc ccctttgttg ttggattatg gagtactaaa
     3781 aattgatttg gtacagcgtc gagtggaata tcagggcaat tttgtcgacc tcacccccca
     3841 ggaattcagt ttgctctacg tgctgaccca agcggaaggt agtgccctca gccgcacaga
     3901 gctactccgc cgggcctggc ccgaggcgat cgacaatccc cgcaccatcg acacccacgt
     3961 cctttccctc cggaaaaaaa ttgagacaga ccccaggcaa cccagtctaa ttcagactgt
     4021 gcgaaatgtg ggctaccgtt ttaactcgtc tattttggaa gagaagaaag ttgtggcgga
     4081 gaaacctgtt accctttccc ccgccggagt ctctgtactt acctaataac aaagcccgaa
     4141 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4201 ctaaacgggt cttgacgggt tttttgtcta gatcaacggc ctcaacctac tactgggctg
     4261 cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc
     4321 gcatttacag ctagctcagt cctaggtaca atgctagccg gaggagggtc aattcatggt
     4381 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4441 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4501 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4561 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4621 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4681 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4741 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4801 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4861 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4921 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4981 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     5041 cgactttagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     5101 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     5161 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     5221 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5281 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5341 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     5401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5461 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccgggatctc
     5521 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggttctcg agcgcatgca
     5581 taaaaactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     5641 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     5701 ggacgcacac cgtggaaacg gatgaaggca cgaacccagt tgacataagc ctgttcggtt
     5761 cgtaaactgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
     5821 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttgta
     5881 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
     5941 ttatggagca gcaacgatgt tacgcagcag caacgatgtt acgcagcagg gcagtcgccc
     6001 taaaacaaag ttaggtggct caagtatggg catcattcgc acatgtaggc tcggccctga
     6061 ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt cgtgagttcg gagacgtagc
     6121 cacctactcc caacatcagc cggactccga ttacctcggg aacttgctcc gtagtaagac
     6181 attcatcgcg cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt
     6241 tctgcccagg tttgagcagc cgcgtagtga gatctatatc tatgatctcg cagtctccgg
     6301 cgagcaccgg aggcagggca ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa
     6361 cgcgcttggt gcttatgtga tctacgtgca agcagattac ggtgacgatc ccgcagtggc
     6421 tctctataca aagttgggca tacgggaaga agtgatgcac tttgatatcg acccaagtac
     6481 cgccacctaa caattcgttc aagccgagat cggcttcccg gccgcggagt tgttcggtaa
     6541 attgtcacaa cgccgccagg tggcactttt cggggaaatg tgcgcgcccg cgttcctgct
     6601 ggcgctgggc ctgtttctgg cgctggactt cccgctgttc cgtcagcagc ttttcgccca
     6661 cggccttgat gatcgcggcg gccttggcct gcatatcccg attcaacggc cccagggcgt
     6721 ccagaacggg cttcaggcgc tcccgaaggt ggatcccccc ctcttgccta cagcatctcc
     6781 cccaggggag aattcttcct gtttcaactc cctctaacgt aaacccattg aatttaaaaa
     6841 agactttatg actgctttac tgctccatga ccaacattat tcccttgatc atgaagcctt
     6901 tctctcaacc ctcagcaaca cagaaaattt actcattatt caagatctag atggcgtttg
     6961 catggggtta gtcaaagacc ccttaacccg caaaattgat cctgactata tccgcgccac
     7021 acgcaagttt agagaccact tctttgtcct caccaacggt gaacatgaag gcagaagggg
     7081 agtaaatcgc atcgttgaac gggcatttcg caatgttgaa gccaaagagg aaacaagcta
     7141 tttacctggt ttagcagcag ggggtgtgca atggcagaca gataatggcc aaatttccca
     7201 tcccggtgtt agccaagcag aactcgattt ccttgccaca gtgccagatt taattggtca
     7261 aagtttagga caatttttta ctaaatatgt tgatattttt cccgctgagc ttcaacctga
     7321 gctgatccat gcttctgtca tgcgccctat agtgagtcgt attacaattc actggccgtc
     7381 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     7441 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     7501 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta a
//
