LOCUS       sequence-417392-        5198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even ORI-marker screen destination vector; contains sfYFP dropout 
            for barcoded-ORI insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 3718 to 3752)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: pGL2_213
FEATURES             Location/Qualifiers
     source          1..5198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(485..579)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(600..1259)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1174..1193
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1260..1362)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1449..1471
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1463..1480
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1464..1480
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1780..1791
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1798..2511
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        3718..3752
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     oriT            4678..4786
                     /note="incP origin of transfer"
     primer_bind     complement(4940..4956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4940..4956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4953..4975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      join(5146..5198,1..336)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaggct
       61 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
      121 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
      181 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
      241 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
      301 gtactatcaa caggttgaac tgctgatctt cagatcggcg cgccggccgg ccgaagatcc
      361 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
      421 gcgtaaatgg gtagggggct tcaaatcgtc cccccatacg atataagttg ttactagtgc
      481 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
      541 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
      601 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
      661 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
      721 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
      781 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
      841 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
      901 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
      961 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     1021 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
     1081 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     1141 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     1201 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     1261 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     1321 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttgatacctg
     1381 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     1441 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     1501 ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg ctattgttgt
     1561 atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgaggtct caggagaatg
     1621 tgaagagctt cgaaagtgaa acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt
     1681 atttaataat gtgtgcggca attcacattt aatttatgaa tgttttctta acatcgcggc
     1741 aactcaagaa acggcaggtt cggatcttag ctactagaga aagaggagaa atactagatg
     1801 cgtaaaggcg aagaactgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
     1861 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
     1921 ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc gactctggta
     1981 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
     2041 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
     2101 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
     2161 cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg
     2221 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
     2281 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
     2341 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
     2401 agctaccaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
     2461 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa ataattcagc
     2521 caaaaaactt aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc
     2581 ggcggttttc ttttctcttc tcaagaagct cttcagcaag gagggccggc ccaatgaata
     2641 ggtttacact tactttagtt ttatggaaat gaaagatcat atcatatata atctagaata
     2701 aaattaacta aaataattat tatctagata aaaaatttag aagccaatga aatctataaa
     2761 taaactaaat taagtttatt taattaacaa ctatggatat aaaataggta ctaatcaaaa
     2821 tagtgaggag gatatataat gttgaataca tacgaacaaa ttaataaagt gaaaaaaata
     2881 cttcggaaac atttaaaaaa taaccttatt ggtacttaca tgtttggatc aggagttgag
     2941 agtggactaa aaccaaatag tgatcttgac tttttagtcg tcgtatctga accattgaca
     3001 gatcaaagta aagaaatact tatacaaaaa attagaccta tttcaaagaa aataggagat
     3061 aaaagcaact tacgatatat tgaattaaca attattattc agcaagaaat ggtaccgtgg
     3121 aatcatcctc ccaaacaaga atttatttat ggagaatggt tacaagagct ttatgaacaa
     3181 ggatacattc ctcagaagga attaaattca gatttaacca taatgcttta ccaagcaaaa
     3241 cgaaaaaata aaagaatata cggaaattat gacttagagg aattactacc tgatattcca
     3301 ttttctgatg tgagacgcgc cattatggat tcgtcagagg aattaataga taattatcag
     3361 gatgatgaaa ccaactctat attaacttta tgccgtatga ttttaactat ggacacgggt
     3421 aaaatcatac caaaagatat tgcgggaaat gcagtggctg aatcttctcc attagaacat
     3481 agggagagaa ttttgttagc agttcgtagt tatcttggag agaatattga atggactaat
     3541 gaaaatgtaa atttaactat aaactattta aataacagat taaaaaaatt ataagcttaa
     3601 aaattgaaaa aatggtggaa acactttttt caattttttt gttttattat ttaatatttg
     3661 ggaaatattc attctaattg gtaatcagat tttagaagtt taaacgctat acaggagttg
     3721 acggctagct cagtcctagg tacagtgcta gctactagga ggaaaaaaat ggtgagtaag
     3781 ggtgaagccg tgattaaaga gttcatgcgc ttcaaagtgc atatggaggg ctcaatgaac
     3841 ggccatgaat ttgaaattga aggtgaagga gaaggccgtc cgtatgaggg cacccagacc
     3901 gccaaactga aagtcacaaa gggtggcccg ctgccgtttt cgtgggatat tttatcccct
     3961 caattcatgt acggcagccg ggcgtttatt aaacacccgg cggatatccc ggattactat
     4021 aagcaatctt ttccagaagg ctttaaatgg gagcgcgtga tgaattttga agacggaggt
     4081 gctgttaccg tgacccagga cacgagctta gaagacggta ccttgatcta taaggtaaag
     4141 ctgcgtggta ccaatttccc gccggatggg ccagttatgc agaaaaagac tatgggctgg
     4201 gaagcatcga cggagcgtct ttaccccgaa gacggcgtcc tgaaaggcga tattaaaatg
     4261 gccctgcgac tgaaagacgg cggtcgctac cttgcagatt tcaaaacaac gtataaagcg
     4321 aaaaaacctg tgcagatgcc cggggcgtac aacgttgatc gcaaactaga tatcacttca
     4381 cacaacgaag attacacggt agtcgaacag tatgaacgta gcgaggggcg ccatagcacc
     4441 ggaggcatgg atgaactcta taaataagct tcgctcgtgc tcgccgcgac cttcctcttc
     4501 atgcacctct tcggcatcgg gctgcacaac gctacagagg agcttttccg ctgcataacc
     4561 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     4621 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     4681 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc
     4741 gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtacg ctaggtatac
     4801 ttgagaccac cccaggatac atagattacc acaactccga gcccttccac ctagcgtgct
     4861 aaccacgcac gctattgttg tataggctag gtggaggctc agtgatgata agtctgcgat
     4921 ggtggatgca tgtgtcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcagag
     4981 ggcacaatcc tattccgcgc tatccgacaa tctccaagac attaggtgga gttcagttcg
     5041 gcgtatggca tatgtcgctg gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5101 ccgtaaaaag gccgcgttgc tggcgttaca gctgggcgcg ccccatgtca gccgttaagt
     5161 gttcctgtgt cactcaaaat tgctttgaga ggctctaa
//
