LOCUS       Exported                5928 bp ds-DNA     circular SYN 16-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5928)
  AUTHORS   Angelica Castaneda
  TITLE     Direct Submission
  JOURNAL   Exported Nov 20, 2018 from SnapGene 4.2.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5928
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    820..838
                     /label=Tet operator 2
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    841..859
                     /label=Tet operator 2(1)
     misc_feature    923..944
                     /label=Primer 178
     primer_bind     1017..1055
                     /label=Fragment.FOR
     CDS             1032..1802
                     /codon_start=1
                     /label=ORF18 Coding Region
                     /translation="MLGKYVCETEPLSPGLRRLMWRFLQNKNLNTFHAQELRFIHLVLC
                     RMYNFGLNVYLLREATANAGTYDEVVLGRKVPAEVWKLVYDGLEEMGVSSEMLLCEAYR
                     DSLWMHLNDKVGLLRGLANYLFHRLGVTHDVRIAPENLVDGNFLFNLGSVLPCRLLLAA
                     GYCLAFWGSDEHERWVRFFAQKLFICYLIVSGRLMTQRSLLVWASETGYPGPVEAVCRD
                     IRSMYGIRTYAVSGYLPAPSEAQLAYLGAFNNNAV"
     primer_bind     complement(1783..1821)
                     /label=Fragment.REV
     CDS             1848..1913
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1923..1925
                     /label=Stop codon
     polyA_signal    1945..2169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2008..2026
                     /label=Primer 179
     rep_origin      2215..2643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2232..2538
                     /label=F1 ori
     promoter        2657..2986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2837..2972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3034..3081
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3100..3474
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3604..3725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3770..3790)
                     /label=M13-rev
     primer_bind     complement(3774..3790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(3796..3818)
                     /label=LacO
     protein_bind    3798..3814
                     /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)."
     promoter        complement(3822..3852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3867..3888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4176..4761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4932..5792)
                     /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"
     CDS             complement(4935..5594)
                     /label=AmpR
     promoter        complement(5793..5897)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatgctcgga aaatacgtgt gtgagaccga gccgctgagc ccggggctta
     1081 ggaggctcat gtggcgcttt ttgcaaaata agaatttaaa tacattccac gcccaagagc
     1141 tgcgttttat tcatttggtt ctctgcagga tgtacaattt cggtctaaat gtgtacctgt
     1201 taagggaggc tactgccaat gccgggacct acgacgaggt ggtcctggga cgcaaggttc
     1261 ctgcggaggt gtggaagctc gtgtacgatg ggctcgagga gatgggcgtg tcaagtgaga
     1321 tgctgctgtg tgaggcatac cgggacagcc tctggatgca cttgaacgat aaggtggggc
     1381 tcttgagggg cctggcgaat tatctgtttc accggctagg ggtcacccac gacgttcgca
     1441 tcgccccgga aaacctggtg gacggaaact ttttgtttaa tctgggaagt gtgctcccct
     1501 gcaggctgct ccttgcggcg ggctactgcc tcgccttttg gggcagcgat gaacacgaac
     1561 gctgggtgcg cttcttcgcc cagaagcttt tcatttgcta cctgatagtc tccgggcgtc
     1621 ttatgacaca gaggtctctg ctagtttggg ccagcgaaac gggctatccc ggtccggtgg
     1681 aggcagtctg tcgcgacatc cgctccatgt acggcatacg aacgtatgcg gtctcgggtt
     1741 atcttccggc tccgtccgaa gcgcagctgg cctaccttgg tgcgtttaac aacaacgcgg
     1801 tttcagatcc agcacagtgg cggccgctcg agggaggcgg tggagccgac tacaaggacc
     1861 acgacggcga ctacaaggac cacgacatcg actacaagga cgacgacgac aaggggcccg
     1921 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1981 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2041 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2101 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2161 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2221 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2281 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2341 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2401 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2461 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2521 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2581 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2641 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2701 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2761 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2821 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2881 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2941 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3001 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     3061 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     3121 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     3181 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     3241 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     3301 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     3361 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     3421 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     3481 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3721 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3781 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4681 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     4741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
