LOCUS Exported 4311 bp ds-DNA circular SYN 12-AUG-2016
KEYWORDS pSPORT2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4311)
AUTHORS Biofeng Lab.
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4311
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 140..156
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 164..182
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature 191..289
/note="MCS"
promoter complement(303..321)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(340..356)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 364..380
/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)."
promoter complement(388..418)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 433..454
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(467..1549)
/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
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(1550..1627)
/gene="lacI"
/note="lacI promoter"
rep_origin complement(1905..2493)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2571..2604
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(2706..3566)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(3854..4309)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(4092..344)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSSNTTHYRESWYACRYRSGIPGAAAVTSELVDSRGSKLT
YACMRRHSSSIVSPKFNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRP
SQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAGFPRQ
ALNRGLPLGFRFSALRHLDPKKLD"
ORIGIN
1 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
61 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
121 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgaatttagg tgacactata
181 gaagagctat gacgtcgcat gcacgcgtac gtaagcttgg atcctctaga gtcgacgagc
241 tcactagtga cagcggccgc cccgggaatt ccggaccggt acctgcaggc gtaccagctt
301 tccctatagt gagtcgtatt agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
541 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
601 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
661 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
721 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
781 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
841 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
901 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
961 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
1021 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
1081 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
1141 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacccgtt
1201 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
1261 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
1321 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
1381 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
1441 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
1501 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
1561 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
1621 tggtgtcaac gtaaatgccg cttcgccttc gcgcgcgaat tgcaagctct gcattaatga
1681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
2101 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
2341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgcc ataacttcgt
2581 atagcataca ttatacgaag ttatggcatg agattatcaa aaaggatctt cacctagatc
2641 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
2701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
2761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
2821 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
2881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
2941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
3001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
3061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
3121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
3181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
3241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
3301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
3361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
3421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
3481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
3541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
3601 cagggttatt gtctcatgcc aggggtgggc acacatattt gataccagcg atccctacac
3661 agcacataat tcaatgcgac ttccctctat cgcacatctt agacctttat tctccctcca
3721 gcacacatcg aagctgccga gcaagccgtt ctcaccagtc caagacctgg catgagcgga
3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
3841 aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
3901 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
3961 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
4021 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
4081 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
4141 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
4201 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
4261 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc c
//