LOCUS       Exported                6029 bp ds-DNA     circular SYN 14-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6029)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6029
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(347..366)
                     /label=oJDW 249
     primer_bind     complement(347..365)
                     /label=oJDW249 AmpR Rev
     primer_bind     882..901
                     /label=oJDW856 AmpR Fwd
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1735..1757
                     /label=Primer 1976
     primer_bind     1921..1938
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1987..2049)
                     /label=PiggyBac 3' ITR
     primer_bind     2227..2260
                     /label=oJDW 1372 attR4 + SpeI
     protein_bind    2251..2375
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2416..2721)
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    complement(2495..2515)
                     /label=RFA Seq Primer 2
     primer_bind     complement(2495..2515)
                     /label=oJDW 93
     CDS             complement(3041..3721)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(3109..3129)
                     /label=oJDW 1396
     primer_bind     3636..3655
                     /label=oJDW 258
     primer_bind     complement(3669..3693)
                     /label=oJDW 1397
     misc_feature    complement(3773..3793)
                     /label=RFA Seq Primer 1
     primer_bind     3773..3793
                     /label=oJDW 92
     promoter        complement(3775..3805)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3831..3954)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    complement(3962..3978)
                     /label=M13 For -20
     primer_bind     complement(3962..3978)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3969..3990)
                     
     primer_bind     complement(3987..4003)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3988..4004)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        4024..4340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4191..4326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4253..4272
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4388..4435
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4454..5053
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5093..5112)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5137..5185
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5401..5422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   5706..5740
                     /label=piggyBac 5' ITR
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     5920..5939
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgct gtacagctta agtcatgttc tttcctgcgt tatcccctga ttctgtggat
     1861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1921 agcgagtcag tgagcgagga agctgtaaaa cgacggccag tagctccacc gccgcagcta
     1981 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat
     2041 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt
     2101 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg
     2161 tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
     2221 tttatgacta gtcatgatta cgccaagcta tcaactttgt atagaaaagt tgaacgagaa
     2281 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     2341 taatactgta aaacacaaca tatccagtca ctatggtcga cctgcagact ggctgtgtat
     2401 aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt
     2461 ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca cactggccat
     2521 atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg
     2581 ggggacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg
     2641 cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct ctcttttata
     2701 ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcggcaaag gagccgttca
     2761 tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg
     2821 cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cgaaccggag atattgacat
     2881 catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt
     2941 catagcatac ctctttttga catacttcgg gtatacatat cagtatatat tcttataccg
     3001 caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatcctcta
     3061 gattacgccc cgcctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     3121 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     3181 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     3241 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     3301 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     3361 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     3421 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     3481 caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt
     3541 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     3601 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     3661 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     3721 ttttagcttc cttagctcct gaaaatctcg acggatccta actcaaaatc cacacattat
     3781 acgagccgga agcataaagt gtaaagcctg ggggtgccta atgcggccgc catagtgact
     3841 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     3901 tattgatatt tatatcattt tacgtttctc gttcaacttt attatacata gttgataatt
     3961 cactggccgt cgttttacgg taccatcgat accgtcgacc tcgacatggt catagctgtt
     4021 gacggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4381 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4441 gaggaactaa accatgaccg agtacaagcc taccgtgcgc ctggccactc gcgatgatgt
     4501 gccccgcgcc gtccgcactc tggccgccgc tttcgccgac taccccgcta cccggcacac
     4561 cgtggacccc gaccggcaca tcgagcgtgt gacagagttg caggagctgt tcctgacccg
     4621 cgtcgggctg gacatcggca aggtgtgggt agccgacgac ggcgcggccg tggccgtgtg
     4681 gactaccccc gagagcgttg aggccggcgc cgtgttcgcc gagatcggcc cccgaatggc
     4741 cgagctgagc ggcagccgcc tggccgccca gcagcaaatg gagggcctgc ttgcccccca
     4801 tcgtcccaag gagcctgcct ggtttctggc cactgtagga gtgagccccg accaccaggg
     4861 caagggcttg ggcagcgccg tcgtgttgcc cggcgtagag gccgccgaac gcgccggtgt
     4921 gcccgccttt ctcgaaacaa gcgcaccaag aaaccttcca ttctacgagc gcctgggctt
     4981 caccgtgacc gccgatgtcg aggtgcccga gggacctagg acctggtgta tgacacgaaa
     5041 acctggcgcc taacgtaccg gttagtaatg agtttaaacg ggggaggcta actgaaacac
     5101 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5161 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5221 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5281 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5341 cctgccatag cagatccggt taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5401 ccagtttgga acaagagtcc acgctgtccc tgatatctat aacaagaaaa tatatatata
     5461 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
     5521 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
     5581 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
     5641 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
     5701 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaatctagc tgcatcaagg
     5761 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5821 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5881 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5941 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6001 gcttaatgcg ccgctacagg gcgcgtcag
//
