LOCUS       Exported                5798 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCSDest2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  AUTHORS   Villefranc JA, Amigo J, Lawson ND
  TITLE     Gateway compatible vectors for analysis of gene function in the 
            zebrafish.
  JOURNAL   Dev Dyn. 2007 Nov . 236(11):3077-87.
  PUBMED    17948311
REFERENCE   2  (bases 1 to 5798)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5798
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    108..232
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        257..287
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             341..1000
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(407..426)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1342..1647
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1526..1545
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1684..1807)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1825..1842)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1847..1981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1871..1890)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1925..1944
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2094..2114)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2094..2112)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2133..2149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2133..2149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2146..2168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2157..2173
                     /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(2181..2211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2226..2247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2364..2381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2535..3123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2615..2634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3294..4154)
                     /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"
     primer_bind     3917..3936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4155..4259)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4285..4740)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4422..4443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4634..4653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4814..5798
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
      181 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcatat tggcggccgc
      241 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
      301 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
      361 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
      421 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
      481 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
      541 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
      601 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
      661 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
      721 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
      781 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
      841 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
      901 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
      961 attacaacag tactgcgatg agtggcaggg cggggcgtaa agatctggat ccggcttact
     1021 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     1081 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     1141 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     1201 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     1261 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     1321 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     1381 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     1441 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     1501 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     1561 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     1621 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     1681 gtccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
     1741 atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac tttattatac
     1801 atagttgcac tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1861 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1921 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1981 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2041 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     2101 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2941 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4261 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4321 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4381 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4441 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4501 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4561 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4621 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4681 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4741 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4801 tattacgcca gtcgaccata gccaattcaa tatggcgtat atggactcat gccaattcaa
     4861 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4921 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4981 tatggcggac ttggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     5041 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     5101 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     5161 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     5221 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     5281 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     5341 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     5401 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     5461 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     5521 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     5581 taacttggca agtacattac tattggaagg acgccagggt acattggcag tactcccatt
     5641 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     5701 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     5761 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
