LOCUS       Exported                5313 bp ds-DNA     circular SYN 01-JUL-2003
DEFINITION  pCDFDuet-1 (Strep).
ACCESSION   .
VERSION     .
KEYWORDS    pCDFDuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 11, 2021 from SnapGene Viewer 4.0.6
            http://www.snapgene.com
COMMENT     ORIGIN
FEATURES             Location/Qualifiers
     source          1..5313
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pGV3319"
     misc_feature    1..19
                     /label=ACYCUP1
                     /note="ACYCUP1"
     promoter        44..60
                     /label=T7 Promoter
                     /note="/dnas_title=T7 Promoter
                     /vntifkey=30"
     protein_bind    65..85
                     /label=lacO
                     /note="/dnas_title=lacO
                     /vntifkey=31"
     RBS             118..123
                     /label=RBS-1
                     /note="/dnas_title=RBS-1
                     /vntifkey=32"
     misc_binding    143..160
                     /label=His-Tag
                     /note="/dnas_title=His-Tag
                     /vntifkey=20"
     CDS             170..1939
                     /codon_start=1
                     /label=Bst10 PolA LF
                     /note="Bst10 PolA LF"
                     /translation="PSSEEEKPLAKMAFTLADRVTEEMLADKAALVVEVVEENYHDAPI
                     VGIAVVNEHGRFFLRPETALADPQFVAWLGDETKKKSMFDSKRAAVALKWKGIELCGVS
                     FDLLLAAYLLDPAQGVDDVAAAAKMKQYEAVRPDEAVYGKGAKRAVPDEPVLAEHLVRK
                     AAAIWALERPFLDELRRNEQDRLLVELEQPLSSILAEMEFAGVKVDTKRLEQMGEELAE
                     QLRTVEQRIYELAGQEFNINSPKQLGVILFEKLQLPVLKKTKTGYSTSADVLEKLAPYH
                     EIVENILHYRQLGKLQSTYIEGLLKVVRPDTKKVHTIFNQALTQTGRLSSTEPNLQNIP
                     IRLEEGRKIRQAFVPSESDWLIFAADYSQIELRVLAHIAEDDNLMEAFRRDLDIHTKTA
                     MDIFQVSEDEVTPNMRRQAKAVNFGIVYGISDYGLAQNLNISRKEAAEFIERYFESFPG
                     VKRYMENIVQEAKQKGYVTTLLHRRRYLPDITSRNFNVRSFAERMAMNTPIQGSAADII
                     KKAMIDLNARLKEERLQARLLLQVHDELILEAPKEEMERLCRLVPEVMEQAVTLRVPLK
                     VDYHYGSTWYDAK"
     CDS             172..2020
                     /codon_start=1
                     /label=MCS-2 ORF
                     /note="/dnas_title=MCS-2 ORF
                     /vntifkey=4"
                     /translation="VIRRGKTACQDGIYAC*PRDGGDACRQGGACR*SGRGKLS*CADR
                     RHRCGQRTWTVFPAPGDGACRSAVCRLAW**NEEKKHV*LKARGSRLEMERN*AMRRFL
                     *FIAGRLFA*SGARC**CGCRSKNEAIRSGAPG*SGVWQRGEAGRAG*ASARRAFGPQG
                     GGDLGARTAVFG*AAPQRTRSVARRARAAVVFDFGGNGICRSESGYEAARTDGRRARRA
                     AAHGRAAHL*ARRPRIQHQFTETARRHFI*KTAAARLEKNENRLLHFGGCA*KTCALSR
                     DRGKHFALPPAWQVAVDVY*RIAESRATRYKEGAYDFQSGVDANRTAQLDGAELAKHSD
                     SA*GRTENPPSVRAIGV*LAHFRRRLLAN*VARPRPYCGR*QFNGSVPPRFGYPYENSD
                     GHFPSERGRSDAQHAPSGEGGQLWDRLRDQ*LRLGAKLKYFAQRGG*IHRALLRKLPWR
                     EAVYGKHCARSKTERVCDDAAASAPLFAGYHEPQLQRPQLC*TDGDEHADSRERR*HY*
                     KGDDRSERQTEGRAAASAPFAAGA*RAHFGGAERRDGAAVPARSGSDGASGHTSRAAQS
                     RLPLWLDVV*CEIRYPRVW*RNRCCEI*TPAHGLVY*RSL"
     protein_bind    1958..2002
                     /label=S-Tag
                     /note="/dnas_title=S-Tag
                     /vntifkey=31"
     primer_bind     complement(2040..2058)
                     /label=T7 Terminator primer
     terminator      2054..2101
                     /label=T7 Terminator
                     /note="/dnas_title=T7 Terminator
                     /vntifkey=43"
     CDS             complement(2275..3063)
                     /codon_start=1
                     /label=aadA (Correct, original source Tn21)
                     /note="/dnas_title=aadA (Correct, original source Tn21)
                     /vntifkey=4"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     RBS             complement(3069..3073)
                     /label=RBS
                     /note="/dnas_title=RBS
                     /vntifkey=32"
     -10_signal      complement(3105..3111)
                     /label=-10 Ap
                     /note="/dnas_title=-10 Ap
                     /vntifkey=47"
     -35_signal      complement(3128..3133)
                     /label=-35 Ap
                     /note="/dnas_title=-35 Ap
                     /vntifkey=48"
     rep_origin      complement(3203..3941)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="/dnas_title=CloDF13 ori
                     /vntifkey=33"
     rep_origin      3294
                     /label=Replication Origin
                     /note="/dnas_title=Replication Origin
                     /vntifkey=33"
     promoter        3746..3775
                     /label=RNA I promoter
                     /note="/dnas_title=RNA I promoter
                     /vntifkey=30"
     promoter        3746..3751
                     /label=Promoter_P_5
                     /note="-35 region (RNA I)
                     /dnas_title=Promoter_P_5
                     /vntifkey=30"
     promoter        3770..3775
                     /label=Promoter_P_6
                     /note="-10 region (RNA I)
                     /dnas_title=Promoter_P_6
                     /vntifkey=30"
     terminator      complement(3858..3879)
                     /label=Terminator_1
                     /note="put. stem-loop structure; put. transcription
                     terminator (RNA I)
                     /dnas_title=Terminator_1
                     /vntifkey=43"
     repeat_region   3858..3866
                     /label=Repeat_Unit_1
                     /label=nonstandard type: repeat_unit
                     /note="inverted repeat A'
                     /dnas_title=Repeat_Unit_1
                     /vntifkey=35"
     repeat_region   3871..3879
                     /label=Repeat_Unit_2
                     /label=nonstandard type: repeat_unit
                     /note="inverted repeat A
                     /dnas_title=Repeat_Unit_2
                     /vntifkey=35"
     promoter        complement(3898..3926)
                     /label=RNA II Promoter
                     /note="/dnas_title=RNA II Promoter
                     /vntifkey=30"
     promoter        complement(3898..3903)
                     /label=Promoter_P_7
                     /note="-10 region (RNA II)
                     /dnas_title=Promoter_P_7
                     /vntifkey=30"
     terminator      complement(3915..3937)
                     /label=Terminator_2
                     /note="put. stem-loop structure; put. transcription
                     terminator (cex gene)
                     /dnas_title=Terminator_2
                     /vntifkey=43"
     repeat_region   3915..3922
                     /label=Repeat_Unit_3
                     /label=nonstandard type: repeat_unit
                     /note="inverted repeat B'
                     /dnas_title=Repeat_Unit_3
                     /vntifkey=35"
     promoter        complement(3921..3926)
                     /label=Promoter_P_8
                     /note="-35 region (RNA II)
                     /dnas_title=Promoter_P_8
                     /vntifkey=30"
     repeat_region   3930..3937
                     /label=Repeat_Unit_4
                     /label=nonstandard type: repeat_unit
                     /note="inverted repeat B
                     /dnas_title=Repeat_Unit_4
                     /vntifkey=35"
     CDS             complement(4151..5233)
                     /codon_start=1
                     /label=lacI
                     /note="Wildtype lacI gene
                     /dnas_title=lacI
                     /vntifkey=lacI"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ggatctcgac gctctccctt atgcgactcc tgcattagga aattaatacg actcactata
       61 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
      121 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgtcatcaga
      181 agaggaaaaa ccgcttgcca agatggcatt tacgcttgct gaccgcgtga cggaggagat
      241 gcttgccgac aaggcggcgc ttgtcgttga agtggtcgag gaaaattatc atgatgcgcc
      301 gatcgtcggc atcgctgtgg tcaacgaaca tggacggttt ttcctgcgcc cggagacggc
      361 gcttgccgat ccgcagtttg tcgcctggct tggtgatgaa acgaagaaaa aaagcatgtt
      421 tgactcaaag cgcgcggcag tcgccttgaa atggaaagga attgagctat gcggcgtttc
      481 ctttgattta ttgctggccg cctatttgct tgatccggcg caaggtgttg atgatgtggc
      541 tgccgcagca aaaatgaagc aatacgaagc ggtgcgcccg gatgaagcgg tgtatggcaa
      601 aggggcgaag cgggccgtgc cggatgagcc agtgctcgcc gagcatttgg tccgcaaggc
      661 ggcggcgatt tgggcgctcg aacggccgtt tttggatgag ctgcgccgca acgaacaaga
      721 tcggttgctc gtcgagctcg agcagccgtt gtcttcgatt ttggcggaaa tggaatttgc
      781 cggagtgaaa gtggatacga agcggctcga acagatgggc gaagagctcg ccgagcagct
      841 gcgcacggtc gagcagcgca tttatgagct cgccggccaa gaattcaaca tcaattcacc
      901 gaaacagctc ggcgtcattt tatttgaaaa actgcagctg cccgtcttga aaaaaacgaa
      961 aaccggctac tccacttcgg cggatgtgct tgaaaaactt gcgccttatc acgagatcgt
     1021 ggaaaacatt ttgcattacc gccagcttgg caagttgcag tcgacgtata ttgaaggatt
     1081 gctgaaagtc gtgcgacccg atacaaagaa ggtgcatacg attttcaatc aggcgttgac
     1141 gcaaaccgga cggctcagct cgacggagcc gaacttgcaa aacattccga ttcggcttga
     1201 ggaaggacgg aaaatccgcc aagcgttcgt gccatcggag tctgattggc tcattttcgc
     1261 cgccgactac tcgcaaattg agttgcgcgt cctcgcccat attgcggaag atgacaattt
     1321 aatggaagcg ttccgccgcg atttggatat ccatacgaaa acagcgatgg acattttcca
     1381 agtgagcgag gacgaagtga cgcccaacat gcgccgtcag gcgaaggcgg tcaactttgg
     1441 gatcgtttac gggatcagtg attacggctt ggcgcaaaac ttaaatattt cgcgcaaaga
     1501 ggcggctgaa ttcatcgagc gctacttcga aagcttccct ggcgtgaagc ggtatatgga
     1561 aaacattgtg caagaagcaa aacagaaagg gtatgtgacg acgctgctgc atcggcgccg
     1621 ctatttgccg gatatcacga gccgcaactt caacgtccgc agctttgctg aacggatggc
     1681 gatgaacacg ccgattcaag ggagcgccgc tgacattatt aaaaaggcga tgatcgatct
     1741 gaacgccaga ctgaaggaag agcggctgca agcgcgcctt ttgctgcagg tgcatgacga
     1801 gctcattttg gaggcgccga aagaagagat ggagcggctg tgccggctcg ttccggaagt
     1861 gatggagcaa gcggtcacac ttcgcgtgcc gctcaaagtc gattaccatt atggctcgac
     1921 gtggtatgat gcgaaataag gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
     1981 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     2041 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2101 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     2161 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg
     2221 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc
     2281 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     2341 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     2401 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     2461 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     2521 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     2581 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     2641 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     2701 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     2761 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     2821 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     2881 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     2941 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     3001 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     3061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3121 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc
     3181 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt
     3241 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc
     3301 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg
     3361 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag
     3421 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt
     3481 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca
     3541 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac
     3601 tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg
     3661 aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt
     3721 tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag
     3781 gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac
     3841 tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga
     3901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct
     3961 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     4021 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4081 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     4141 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4261 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4321 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4381 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     4441 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4501 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4561 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4621 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4681 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4741 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4801 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4861 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4921 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4981 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     5041 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     5101 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     5161 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     5221 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5281 cgcgaaaggt tttgcgccat tcgatggtgt ccg
//
