LOCUS       Exported                5763 bp ds-DNA     circular SYN 09-MAY-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 2019 from SnapGene Viewer 4.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5763
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    89..511
                     /label=ZF_ADD_GCN
     CDS             437..493
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     terminator      575..622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      659..1114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1141..1245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1246..2106
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2277..2865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3051..3193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3295..3486)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4295..5377)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5378..5455)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggaacgtccg tacgcttgcc cggttgaatc
      121 ttgcgaccgt cgtttctctg acaaaaccaa actgcgtgtt catattagaa ttcatactgg
      181 acaaaaacca ttccaatgta gaatttgtat gagaaatttc tctgttcgtc acaacctgac
      241 ccgtcacatc cgtacccaca ccggtgaaaa accgttcgct tgcgacatct gcggtcgtaa
      301 attcgctcgt tctgacgaac gtaaacgtca caccaaaatc cacctgcgtc agaaagacgg
      361 tggcggcggt tctccagcgc cagcgccacc gcatcgcgat attcagcata ttctgccgat
      421 tctggaagat aaagtggaag aactgctgag caaaaactat catctggaaa acgaagtggc
      481 gcgcctgaaa aaactggtgg gcgaacgcta agatccggct gctaacaaag cccgaaagga
      541 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      601 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      721 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      781 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      841 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      901 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      961 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1021 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1081 gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg
     1141 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1201 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1261 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1321 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1381 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1441 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1501 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1561 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1621 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1681 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1741 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     1801 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1861 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1921 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1981 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2041 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2101 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2161 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2221 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2281 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2341 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2401 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2461 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2521 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2581 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2641 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2701 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2761 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2821 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2881 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2941 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3001 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3061 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     3121 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3181 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3241 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3301 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3361 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3421 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3481 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3541 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3601 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3661 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3721 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3781 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3841 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3901 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     3961 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     4021 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     4081 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4141 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4201 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4261 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4321 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4381 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     4441 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4501 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4561 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4621 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4681 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4741 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4801 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4861 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4921 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4981 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     5041 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     5101 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     5161 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     5221 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     5281 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     5341 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     5401 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     5461 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     5521 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     5581 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     5641 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     5701 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     5761 aat
//
