LOCUS Exported File 4930 bp ds-DNA circular SYN 03-JAN-2013

DEFINITION .

ACCESSION .

VERSION .

KEYWORDS pTCP (empty vector)

SOURCE synthetic DNA construct

ORGANISM synthetic DNA construct

REFERENCE 1 (bases 1 to 4930)

AUTHORS Trial User

TITLE Direct Submission

JOURNAL Exported Thursday, Jan 03, 2013 from SnapGene 1.4.0

FEATURES Location/Qualifiers

source 1..4930

/organism="synthetic DNA construct"

/mol_type="other DNA"

promoter 365..568

/note="CMV promoter"

/note="human cytomegalovirus (CMV) immediate early

promoter"

/note="color: #ffffff; direction: RIGHT"

misc_feature 652..661

/note="Site of cDNA insert"

/note="color: #ff99cc"

rep_origin 815..1243

/direction=RIGHT

/note="f1 ori"

/note="f1 bacteriophage origin of replication; arrow

indicates direction of (+) strand synthesis"

/note="color: #ffff00"

promoter 1257..1586

/note="SV40 promoter"

/note="SV40 enhancer and early promoter"

/note="color: #ffffff; direction: RIGHT"

CDS 1598..2197

/codon_start=1

/gene="pac from Streptomyces alboniger"

/product="puromycin N-acetyltransferase"

/note="PuroR"

/note="confers resistance to puromycin"

/note="color: #ccffcc"

/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER

VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA

AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS

APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"

polyA_signal 2356..2477

/note="SV40 poly(A) signal"

/note="SV40 polyadenylation signal"

/note="color: #a6acb3"

primer_bind complement(2526..2542)

/note="M13 rev"

/note="common sequencing primer, one of multiple similar

variants"

/note="color: #a020f0; direction: LEFT"

rep_origin complement(2928..3513)

/direction=LEFT

/note="ori"

/note="high-copy-number colE1/pMB1/pBR322/pUC origin of

replication"

/note="color: #ffff00"

CDS complement(3684..4544)

/codon_start=1

/gene="bla"

/product="beta-lactamase"

/note="AmpR"

/note="confers resistance to ampicillin, carbenicillin, and

related antibiotics"

/note="This feature has 2 segments:

1:3684..4475/#ccffcc

2:4476..4544/#ccffcc/signalsequence

Cleavage site after base 4475"

/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

LIKHW"

promoter complement(4545..4649)

/gene="bla"

/note="AmpR promoter"

/note="color: #ffffff; direction: LEFT"

enhancer 4915..364

/note="CMV enhancer"

/note="human cytomegalovirus immediate early enhancer"

/note="color: #ffffff"

ORIGIN

1 tagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccg

61 cgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccatt

121 gacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtca

181 atgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgcc

241 aagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagta

301 catgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattac

361 catggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacgggg

421 atttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacg

481 ggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgt

541 acggtgggaggtctatataagcagagctggtttagtgaaccgtggatcccgtcgcttacc

601 gattcagaatggttgatatccgccattctgaatcggtaagcgacgaagcttaataaagga

661 tcttttattttcattggatctgtgtgttggttttttgtgtgcggccgccctcgactgtgc

721 cttctagaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcg

781 gaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcattaagc

841 gcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgccc

901 gctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagct

961 ctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaa

1021 aaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgc

1081 cctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaaca

1141 ctcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctat

1201 tggttaaaaaatgagctgatttaacaaaaatttaacgcgaattaattctgtggaatgtgt

1261 gtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgc

1321 atctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagta

1381 tgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcc

1441 cgcccctaactccgcccagttccgcccattctccgccccatggctgactaatttttttta

1501 tttatgcagaggccgaggccgcctctgcctctgagctattccagaagtagtgaggaggct

1561 tttttggaggcctaggcttttgcaaaaagctcccgggatgaccgagtacaagcccacggt

1621 gcgcctcgccacccgcgacgacgtcccgcgggccgtacgcaccctcgccgccgcgttcgc

1681 cgactaccccgccacgcgccacaccgtcgacccggaccgccacatcgagcgggtcaccga

1741 gctgcaagaactcttcctcacgcgcgtcgggctcgacatcggcaaggtgtgggtcgcgga

1801 cgacggcgccgcggtggcggtctggaccacgccggagagcgtcgaagcgggggcggtgtt

1861 cgccgagatcggcccgcgcatggccgagttgagcggttcccggctggccgcgcagcaaca

1921 gatggaaggcctcctggcgccgcaccggcccaaggagcccgcgtggttcctggccaccgt

1981 cggcgtctcgcccgaccaccagggcaagggtctgggcagcgccgtcgtgctccccggagt

2041 ggaggcggccgagcgcgccggggtgcccgccttcctggagacctccgcgccccgcaacct

2101 ccccttctacgagcggctcggcttcaccgtcaccgccgacgtcgaggtgcccgaaggacc

2161 gcgcacctggtgcatgacccgcaagcccggtgcctgattcgaatgaccgaccaagcgacg

2221 cccaacctgccatcacgagatttcgattccaccgccgccttctatgaaaggttgggcttc

2281 ggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggag

2341 ttcttcgcccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagc

2401 atcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaa

2461 ctcatcaatgtatcttatcatgtctgtataccgtcgacctctagctagagcttggcgtaa

2521 tcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacata

2581 cgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacatta

2641 attgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaa

2701 tgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcg

2761 ctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaag

2821 gcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaa

2881 ggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctc

2941 cgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgaca

3001 ggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccg

3061 accctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttct

3121 catagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgt

3181 gtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgag

3241 tccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagc

3301 agagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctac

3361 actagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaaga

3421 gttggtagctcttgatccggcaaacaaaccaccgctggtagcggtttttttgtttgcaag

3481 cagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacgggg

3541 tctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaa

3601 aggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtata

3661 tatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcg

3721 atctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgata

3781 cgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccg

3841 gctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcct

3901 gcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagt

3961 tcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgc

4021 tcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatga

4081 tcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagt

4141 aagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtc

4201 atgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaa

4261 tagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgcca

4321 catagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctca

4381 aggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatct

4441 tcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgcc

4501 gcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaa

4561 tattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatt

4621 tagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtc

4681 gacggatcgggagatctcccgatcccctatggtgcactctcagtacaatctgctctgatg

4741 ccgcatagttaagccagtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcg

4801 cgagcaaaatttaagctacaacaaggcaaggcttgaccgacaattgcatgaagaatctgc

4861 ttagggttaggcgttttgcgctgcttcgcgatgtacgggccagatatacgcgttgacatt

4921 gattattgac

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