LOCUS       Exported                9059 bp ds-DNA     circular SYN 12-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    TIAM1_pCSdest
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9059)
  AUTHORS   Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF, 
            McCallion AS, Katsanis N, Reeves RH
  TITLE     Survey of Human Chromosome 21 Gene Expression Effects on Early 
            Development in Danio rerio.
  JOURNAL   G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi: 
            10.1534/g3.118.200144.
  PUBMED    29760202
REFERENCE   2  (bases 1 to 9059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9059
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1085..1109
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6021..6045)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6067..6084)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6089..6223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6113..6132)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6167..6186
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6336..6356)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6336..6354)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6375..6391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6375..6391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6388..6410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6399..6415
                     /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(6423..6453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6468..6489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6606..6623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6777..7365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6857..6876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7536..8396)
                     /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     8159..8178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8397..8501)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8527..8982)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8664..8685)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8876..8895
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcaag
     1081 gatcacaagt ttgtacaaaa aagcaggctt taaaggaacc atctgcagaa ttcgccctta
     1141 gcaagttggg ctggggacca ctgttgcctt ttaagaccat aaaaccatgg gaaacgcaga
     1201 aagtcaacat gtagagcacg agttttatgg agaaaagcat gccagcctgg ggcgcaagca
     1261 cacttcccgc tccctgcgcc tctcgcacaa gacgcggagg accaggcacg cttcctcggg
     1321 gaaggtgatc cacaggaact ccgaagtgag cacccgatcc agcagcaccc ccagcatccc
     1381 ccagtccctg gctgaaaatg gcctggagcc cttctcccaa gatggtaccc tagaagactt
     1441 cgggagcccc atctgggtgg accgagtgga catgggcttg agacctgtgt cttacactga
     1501 ctcttctgtc actcccagcg tagacagcag catcgtcctc acagcagcct ctgtgcagag
     1561 catgccagac actgaggaga gcaggcttta cggggatgac gctacatatt tggctgaggg
     1621 aggcaggagg cagcattcct atacatccaa tgggcccact ttcatggaga cggcgagctt
     1681 taagaagaaa cgctccaaat ctgcagacat ctggcgggag gacagcctgg aattctcact
     1741 ctctgatctg agccaagaac atttaacaag caacgaagaa atcttgggtt ccgccgaaga
     1801 gaaggactgc gaggaggctc gggggatgga aacgcgggcg agtccgcggc agctcagcac
     1861 ctgtcagaga gccaattcct tgggtgactt gtatgctcag aaaaactctg gagtgacagc
     1921 aaacgggggg ccggggagca aatttgcagg ctactgtcgg aatttggtgt ctgatattcc
     1981 caatcttgca aaccataaga tgccaccagc tgctgctgaa gagactcctc cgtacagtaa
     2041 ttataacaca cttccctgta ggaaatctca ctgtctctct gaaggtgcca ccaacccaca
     2101 aattagccat agcaacagca tgcaaggcag aagagctaaa acaactcagg atgttaatgc
     2161 aggcgagggc agtgagtttg cagacagtgg gattgaaggg gccactaccg acacggacct
     2221 cctgtccagg cgatctaatg ccaccaactc cagctactca cccaccacag gccgggcctt
     2281 tgtgggcagc gacagcggca gcagctccac cggggatgcg gctcgtcagg gggtgtacga
     2341 gaacttccgg cgggagctgg agatgagcac caccaacagc gagagcctgg aggaggccgg
     2401 ctcggcgcac agcgatgagc agagcagcgg caccctgagc tctccgggcc agtcggacat
     2461 cctgctgacc gccgcacagg gcacggtgcg caaggccggc gccctggccg tcaagaactt
     2521 cctggtgcac aagaagaaca agaaggtgga gtcagccacc cggaggaagt ggaagcacta
     2581 ctgggtgtcc ctgaaaggat gcacgctatt tttctacgag agcgacggca ggtctgggat
     2641 agaccacaac agcatcccca aacacgccgt ctgggtggag aacagcattg tgcaggcggt
     2701 gcctgagcac cccaagaagg actttgtctt ctgcctcagc aattccctgg gtgatgcctt
     2761 cctttttcag accactagcc agacggagct tgaaaactgg atcaccgcca tccactctgc
     2821 ctgcgccact gcggtcgcga ggcaccacca caaggaagac acgctccgac tcctgaaatc
     2881 agagatcaaa aaactggaac agaagattga catggatgaa aagatgaaga aaatgggtga
     2941 aatgcagctg tcttcagtca ctgactcaaa gaaaaagaaa acaatattag atcagatctt
     3001 tgtctgggag caaaatctcg agcagttcca aatggacctg tttcgtttcc gctgttattt
     3061 agccagcctt cagggtgggg agctgccaaa ccccaaaagg cttctcgctt ttgcaagtcg
     3121 accaacgaaa gtggccatgg gccgccttgg aatcttttcg gtatcatcgt ttcatgccct
     3181 ggtggcagca cgcactggtg aaactggagt gagaagacgt actcaggcca tgtccagatc
     3241 cgcgagcaag cgaaggagca ggttttcttc tctgtggggt ctggatacta cctccaaaaa
     3301 gaagcaggga cggccaagca tcaatcaggt gtttggagag ggaaccgaag ctgtaaagaa
     3361 atctttagag ggaatatttg atgacattgt tccagatggc aagagggaga aagaagtggt
     3421 cttacctaac gttcaccagc acaaccctga ctgcgacatt tgggtccacg agtatttcac
     3481 tccatcctgg ttctgtctgc ccaataatca gcctgccctg acggtcgtcc ggccaggcga
     3541 cactgcacgg gacaccctgg agctgatttg caagacacat caactggatc attctgctca
     3601 ttacctgcgc ctgaaatttc taatagaaaa caaaatgcag ctctatgttc cacagcccga
     3661 ggaagacatc tatgagctgc tgtacaaaga aattgaaatc tgtccaaaag tcactcagag
     3721 catccacatt gagaagtcag atacagctgc tgatacttac gggttttcac tttcttctgt
     3781 ggaagaagat ggtattcgaa ggctgtacgt gaatagtgtg aaggaaaccg gtttagcttc
     3841 caagaaaggc ctgaaagcag gagatgagat tcttgagatc aataatcgtg ctgctgacgc
     3901 cctgaactct tctatgctca aagatttcct ctcacagccc tcgctgggcc tcctggtgag
     3961 gacctacccc gagctggagg aaggagtgga gctgctggaa agcccgcccc accgagtgga
     4021 cggccctgcc gaccttggcg agagccccct cgcctttctc accagcaacc cagggcacag
     4081 cctttgcagc gagcagggca gcagtgctga gaccgctcca gaggagaccg aggggccaga
     4141 cttggaatcc tcagatgaga ctgatcacag cagcaagagt acagaacagg tggccgcatt
     4201 ttgccgcagt ttgcatgaga tgaacccctc tgaccagagc ccatctcctc aggactccac
     4261 ggggcctcag ctggcgacca tgagacaact ctcggatgca gataagctgc gcaaggtgat
     4321 ctgcgagctc ctggagacgg agcgcaccta cgtgaaggat ttaaactgtc ttatggagag
     4381 atacctaaag cctcttcaaa aagaaacttt tctcacccag gatgagcttg acgtgctttt
     4441 tggaaattta acggaaatgg tagagtttca agtagaattc cttaaaactc tagaagatgg
     4501 agtgagactg gtacctgatt tggaaaagct tgagaaggtt gatcaattta agaaagtgct
     4561 gttctctctg gggggatcat tcctgtatta tgctgaccgc ttcaagctct acagtgcctt
     4621 ctgcgccagc cacacaaaag ttcccaaggt cctggtgaaa gccaagacag acacggcttt
     4681 caaggcattc ttggatgccc agaacccgaa gcagcagcac tcatccacgc tggagtcgta
     4741 cctcatcaag cccatccaga ggatcctcaa gtacccactt ctgctcaggg agctgttcgc
     4801 cctgaccgat gcggagagcg aggagcacta ccacctggac gtggccatca agaccatgaa
     4861 caaggttgcc agtcacatca atgagatgca gaaaatccat gaagagtttg gggctgtgtt
     4921 tgaccagctg attgctgaac agactggtga gaaaaaagag gttgcagatc tgagcatggg
     4981 agacctgctt ttgcacacta ccgtgatctg gctgaacccg ccggcctcgc tgggcaagtg
     5041 gaaaaaggaa ccagagttgg cagcattcgt cttcaaaact gctgtggtcc ttgtgtataa
     5101 agatggttcc aaacagaaga agaaacttgt aggatctcac aggctttcca tttatgagga
     5161 ctgggacccc ttcagatttc gacacatgat ccccacggaa gcgctgcagg ttcgagcttt
     5221 ggcgagtgca gatgcagagg caaatgccgt gtgtgaaatt gtccatgtaa aatccgagtc
     5281 tgaagggagg ccggagaggg tctttcactt gtgctgcagc tccccagaga gccgaaagga
     5341 tttcctaaag gctgtgcatt caatcctgcg tgataagcac agaagacagc tcctcaaaac
     5401 cgagagcctt ccctcatccc agcaatatgt cccttttgga ggcaaaagat tgtgtgcact
     5461 gaagggggcc aggccggcca tgagcagggc agtgtctgcc ccaagcaagt ctcttgggag
     5521 gaggaggcgg cggctggctc gaaacaggtt taccattgat tctgatgccg tctccgcaag
     5581 cagcccggag aaagagtccc agcagccccc cggtggtggg gacactgacc gatgggtaga
     5641 ggagcagttt gatcttgctc agtatgagga gcaagatgac atcaaggaga cagacatcct
     5701 cagtgacgat gatgagttct gtgagtccgt gaagggtgcc tcagtggaca gagacctgca
     5761 ggagcggctt caggccacct ccatcagtca gcgggaaaga ggccggaaaa ccctggatag
     5821 tcacgcgtcc cgcatggcac agctcaagaa gcaagctgcc ctgtcgggga tcaatggagg
     5881 cctggagagc gcaagcgagg aagtcatttg ggttaggcgt gaagactttg ccccctccag
     5941 gaaactgaac actgagatct gactgcgtca cctgccccgt agacaagggc gaattccagc
     6001 acactggcgg ccgttactag acccagcttt cttgtacaaa gtggtgatcc tctcgagcct
     6061 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     6121 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     6181 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     6241 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     6301 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     6361 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6421 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8521 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     8581 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     8641 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     8701 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     8761 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     8821 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     8881 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     8941 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     9001 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacc
//
