LOCUS       Exported                6309 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    453 pCSMTN5ICD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6309)
  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 6309)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6309
                     /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"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             322..351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    379..403
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2290..2314)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2336..2353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2358..2492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2382..2401)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2436..2455
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2605..2625)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2605..2623)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2644..2660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2644..2660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2657..2679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2668..2684
                     /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(2692..2722)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2737..2758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2875..2892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3046..3634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3126..3145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3805..4665)
                     /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     4428..4447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4666..4770)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4796..5251)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4933..4954)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5145..5164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5325..6309
                     /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 tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggagcaaaag ctcatttctg aagaggactt gaatttagag
      361 atctcgaatt caaggatcac aagtttgtac aaaaaagcag gcttgatggt tggcatgttg
      421 attgcccgcc gcaagcgtga acacagcaca ctctggttcc ctgagggctt cttcctcaaa
      481 aaggaaacaa gcagtaacaa gaaccgcaga gaacctgtgg gccaagattc actgggcatg
      541 aaacacatgc caaagacagt ggaagagtct cttctggctg atcacagtga ccagtggata
      601 gatacagact gcccagaggc taagcgactt aaggtggaag agccaagcat tctgtcagat
      661 ggtgaggatg cagttgacag cagacagtgg acacagcatc acttggcagc agcagatatc
      721 cgcatgccac cttccatggc acttacacca ccacaaggag agtttgacag tgactgcatg
      781 gatgttaatg tcagaggccc tgatggcttc acacctctga tgctggcatc tttctgtgga
      841 ggagggcttg agccagaggt gaccgaagat gatgactcag atgaatcctc agccaacatc
      901 atatctgacc tcatttacca gggagcatca cttgcagctc aaactgaccg cactggagag
      961 actgctctgc acctggctgc acgctatgcc cgggcagatg cagctaaaag actcctggat
     1021 gctggggctg atgcaaatgc acaggataac acaggtcgct cacctttgca tgcagctgtg
     1081 gcagctgatg cacagggagt tttccagatt ctgatcagga accgtgccac agacttggac
     1141 gcgcgcatgt atgatggctc cactgctctg atacttgcag cccgtctggc agtagagggc
     1201 atggtggaag agttgatcac ctgccatgct gatgttaatg ctgttgatga aatcggaaag
     1261 tcagcattgc actgggcagc agcagtcaat aacgttgaag ctactattgc cttgttgaag
     1321 aatggtgcca ataaagatat gcaagacctc aaggaggaga ctccgctgtt cttggctgcc
     1381 agggaaggca gctgtgaggc agtaaaggtg ttgttggctc actttgccaa tagggaaata
     1441 acagatcaca tggacagact ccctcgagat attgcccagg agcgcatgca tcatgacatt
     1501 gtgcagctac tggatgaata taacactgtg cgaagtccac agggtcatgc aggggcaggt
     1561 caccacctct ctggtagcca cacactctca ccgctcatgt gccctcccag caacttcctc
     1621 caagggctga aaagtacccc tcagggtaag aagaaccgtc gcccgggagc taaaggcatt
     1681 ggtggacagc acgcttccgg cctgaaagat tcagccaaag gacgcaacaa acggctgact
     1741 ctagacatgc agagtgcttt gctggatagc tctgtcacgc tctcccctgt tgactcactg
     1801 gattcaccac gtggaggcgc cagtaacgct ggctacgtca ccaatccaac ctcaccagct
     1861 gccatgccct ctcccggcct tttccactcc tctatgtctg tccctaatac ccctatggtg
     1921 cacagcagta tcctggacag caccagccct tttgctgtct ccctggcaca gttgagtgac
     1981 ctgggagatg gtgggctctc aatgcagggg cgtgtgggca tgcaaggagg gggtgtcaac
     2041 ccaggccccc ataactacgt gatgaacgcc ggtcaaatga gccttagtat gggcatggta
     2101 agtccagtaa gcgtgccgtt tgactggcac aaccgcatgc ctccatcttc tcagtgcagt
     2161 cagccaatga tcagcatggt gcatccagta agtagtcaag caggcatgct cccacaaaca
     2221 ccaaccctgc aaagcagcat gctattgcac cagcaggtgt tccgtaatgc ccagcagccc
     2281 atcctgcaga cccagctttc ttgtacaaag tggtgatcct ctcgagcctc tagaactata
     2341 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     2401 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2461 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2521 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     2581 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2641 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2701 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2761 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2821 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2881 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3481 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3781 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3841 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3901 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3961 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4021 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4081 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4141 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4201 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4261 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4321 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4381 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4441 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4501 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4561 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4621 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4681 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4741 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4801 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4861 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4921 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4981 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5041 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5101 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5161 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5221 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5281 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca
     5341 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     5401 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     5461 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg
     5521 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     5581 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     5641 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     5701 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     5761 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     5821 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     5881 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     5941 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     6001 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     6061 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     6121 gacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     6181 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     6241 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     6301 ttagggaac
//
