LOCUS Exported 5790 bp ds-DNA circular SYN 25-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5790
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 19..35
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 78..330
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(349..1068)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
CDS 1855..2880
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
promoter complement(2947..2965)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2986..3002)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3010..3026
/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(3034..3064)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3079..3100
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3388..3976)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4147..5007)
/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"
promoter complement(5008..5112)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5138..5593)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 5735..5751
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 5758..5776
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attctcatgt
61 ttgacagctt atcatcggat ctagtaacat agatgacacc gcgcgcgata atttatccta
121 gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat
181 cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
241 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
301 gaaactttat tgccaaatgt ttgaacgatc tgcagccggg cggccgcttt acttgtacag
361 ctcgtccatg ccgtgagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
421 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
481 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
541 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
601 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
661 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
721 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
781 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
841 ggggtagcgg ctgaagcact gcacgccgta ggtgaaggtg gtcacgaggg tgggccaggg
901 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
961 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
1021 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg cctatattca
1081 ttcattgtca gctatcgctg gcaacaagat ataagcgtgt acttacgatt ctaggaccct
1141 gcgaatagag ttggagttgg ataccgatgg acgattgtta gataggacgg gatgctgata
1201 gtcgaggtaa gttgttgagg gtggtggttt tatcaaggac ggacacctta gccctttata
1261 tacacagaat cttattgcta ggtagccgga cggtgcggat tccaggtatc agtaggccct
1321 ctgaatgtcc atgcaatgat gcccaacaat cgagacgatc ctacttcgga accgactcga
1381 tcctcgtcgg gcctacatgt atgagaccat gtgcacctcc gaagcatcct tacggagagc
1441 ggaattgcgc gtacagaact cctccatgca ttccaatcga taccgtcgac gttaactgat
1501 attgaaggag cattttttgg gcttggctgg agctagtgga ggtcaacaat gaatgcctat
1561 tttggtttag tcgtccaggc ggtgagcaca aaatttgtgt cgtttgacaa gatggttcat
