LOCUS       Exported                8661 bp ds-DNA     circular SYN 02-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8661)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 5, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     UUID: bd87a248-8108-4940-8310-b8e265dfb50c
            Type: Synthetic
            ConfirmedExperimentally: 0
            CreatedBy: Taylor Jones IV
            SequenceClass: UNA
            TransformedInto: Stellara¢
FEATURES             Location/Qualifiers
     source          1..8661
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /locus_tag="CMV enhancer"
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /locus_tag="CMV promoter"
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     regulatory      597..602
                     /locus_tag="Kozak sequence"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
     CDS             603..1319
                     /locus_tag="mGFP"
                     /label=mGFP
                     /note="/vntifkey=4"
     misc_feature    1356..1637
                     /locus_tag="FRB"
                     /label=FRB
     misc_feature    1638..1745
                     /locus_tag="GSV5GS"
                     /label=GSV5GS
     misc_feature    1668..1709
                     /locus_tag="V5 tag"
                     /label=V5 tag
     CDS             1746..2608
                     /codon_start=1
                     /locus_tag="FBP17 truncated"
                     /label=FBP17 truncated
                     /translation="MSWGTELWDQFDNLEKHTQWGIDILEKYIKFVKERTEIELSYAKQ
                     LRNLSKKYQPKKNSKEEEEYKYTSCKAFISNLNEMNDYAGQHEVISENMASQIIVDLAR
                     YVQELKQERKSNFHDGRKAQQHIETCWKQLESSKRRFERDCKEADRAQQYFEKMDADIN
                     VTKADVEKARQQAQIRHQMAEDSKADYSSILQKFNHEQHEYYHTHIPNIFQKIQEMEER
                     RIVRMGESMKTYAEVDRQVIPIIGKCLDGIVKAAESIDQKNDSQLVIEAYKSGFEPPGD
                     IEFEDY"
     polyA_signal    2736..2857
                     /locus_tag="SV40 poly(A) signal(1)"
                     /label=SV40 poly(A) signal
                     /label=SV40 poly(A) signal(1)
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2864..3319)
                     /direction=LEFT
                     /locus_tag="f1 ori(1)"
                     /label=f1 ori
                     /label=f1 ori(1)
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     enhancer        3342..3645
                     /locus_tag="CMV enhancer(1)"
                     /label=CMV enhancer
                     /label=CMV enhancer(1)
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3646..3849
                     /locus_tag="CMV promoter(1)"
                     /label=CMV promoter
                     /label=CMV promoter(1)
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3887..4207
                     /locus_tag="DmrA(1xFKBP)"
                     /label=DmrA(1xFKBP)
     CDS             4250..4570
                     /locus_tag="DmrA(1xFKBP)(1)"
                     /label=DmrA(1xFKBP)
                     /label=DmrA(1xFKBP)(1)
     CDS             4613..4933
                     /locus_tag="DmrA(1xFKBP)(2)"
                     /label=DmrA(1xFKBP)
                     /label=DmrA(1xFKBP)(2)
     CDS             4976..5665
                     /locus_tag="TagBFP"
                     /label=TagBFP
     misc_feature    5666..5713
                     /locus_tag="CaaX"
                     /label=CaaX
     misc_feature    5699..5710
                     /locus_tag="CAAX"
                     /label=CAAX
     misc_feature    5722..5750
                     /locus_tag="MCS(1)"
                     /label=MCS
                     /label=MCS(1)
                     /note="multiple cloning site"
     misc_feature    5751..5787
                     /locus_tag="MCS"
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    5911..6032
                     /locus_tag="SV40 poly(A) signal"
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6039..6050)
                     /direction=LEFT
                     /locus_tag="f1 ori"
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6059..6163
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
     promoter        6165..6522
                     /locus_tag="SV40 promoter"
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6373..6508
                     /direction=BOTH
                     /locus_tag="SV40 ori"
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6557..7351
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /locus_tag="NeoR/KanR"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (GeneticinA(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7583..7630
                     /locus_tag="HSV TK poly(A) signal"
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7959..8547
                     /direction=RIGHT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1021 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1081 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     1321 gcagtagcgg aagcagtggt tcttcaggat catccatgat cctctggcat gagatgtggc
     1381 atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg aaaggcatgt
     1441 ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact ctgaaggaaa
     1501 catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt
     1561 acatgaaatc agggaatgtc aaggacctcc tccaagcctg ggacctctat tatcatgtgt
     1621 tccgacgaat ctcaaagggg tgtggcagcg gaggtagtgg gagcggtggc aagcccatcc
     1681 ccaaccccct gctgggcctg gacagcacca agggctcggg ctcgacctcg ggctcgggca
     1741 ccggtatgag ctggggcacc gagctctggg atcagtttga caacttagaa aaacacacac
     1801 agtggggaat tgatattctt gagaaatata tcaagtttgt gaaagaaagg acagagattg
     1861 aactcagcta tgcaaagcaa ctcaggaatc tttcaaagaa gtaccaacct aaaaagaact
     1921 cgaaggagga agaagaatac aagtatacgt catgtaaagc tttcatttcc aacctgaacg
     1981 aaatgaatga ttacgcaggg cagcatgaag ttatctccga gaacatggca tcacagatca
     2041 ttgtggactt ggcacgctat gttcaggaac tgaaacagga gaggaaatca aactttcacg
     2101 atggccgtaa agcacagcag cacatcgaga cttgctggaa gcagcttgaa tctagtaaaa
     2161 ggcgatttga acgcgattgc aaagaggcgg acagggcgca gcagtacttt gagaaaatgg
     2221 acgctgacat caatgtcaca aaagcggatg ttgaaaaggc ccgacaacaa gctcaaatac
     2281 gtcaccaaat ggcagaggac agcaaagcag attactcatc cattctccag aaattcaacc
     2341 atgagcagca tgaatattac catactcaca tccccaacat cttccagaaa atacaagaga
     2401 tggaggaaag gaggattgtg agaatgggag agtccatgaa gacatatgca gaggttgatc
     2461 ggcaggtgat cccaatcatt gggaagtgcc tggatggaat agtaaaagca gccgaatcaa
     2521 ttgatcagaa aaatgattca cagctggtaa tagaagctta taaatcaggg tttgagcctc
     2581 ctggagacat tgaatttgag gattacactg ggtagcggcc gcgactctag atcataatca
     2641 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2701 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2881 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2941 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3001 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3061 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3121 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3181 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3241 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3301 gcgccgctac agggcgcgtc aggtggcact tttcggggcc gcgttacata acttacggta
     3361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     3841 agcagagctg gtttagtgaa ccgtcagatc cgctagcgcc accatgggag tgcaggtgga
     3901 aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
     3961 ctacaccggg atgcttgaag atggaaagaa atttgattcc tcccgggaca gaaacaagcc
     4021 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
     4081 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
     4141 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
     4201 actggaactg caggatatca agcttgcggt accgcgggcc cgggatccag gagtgcaggt
     4261 ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt
     4321 gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg acagaaacaa
     4381 gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc
     4441 ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc
     4501 cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct
     4561 aaaactggaa ctgcaggata tcaagcttgc ggtaccgcgg gcccgggatc caggagtgca
     4621 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
     4681 ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa
     4741 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     4801 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     4861 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     4921 tctaaaactg gaactgcagg atatcaagct tgcggtaccg cgggcccggg atccaatgag
     4981 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca
     5041 tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     5101 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt
     5161 cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca
     5221 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     5281 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     5341 aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     5401 cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct
     5461 gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa
     5521 acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat
     5581 caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg
     5641 cgacctccct agcaaactgg ggcacggtaa aaagaagaaa aagaagtcaa agacaaagtg
     5701 tgtaattatg tagtgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
     5761 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     5821 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     5881 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5941 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6001 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg gaaatgtgcg
     6061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     6181 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     6241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     6301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     6361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6421 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     6541 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     6601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6721 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6781 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     6901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     7021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     7081 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     7141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     7201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     7261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     7321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     7381 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     7441 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     7501 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     7561 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     7621 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     7681 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     7741 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     7801 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8581 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8641 aaccgtatta ccgccatgca t
//
